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Information on EC 3.1.21.10 - crossover junction endodeoxyribonuclease
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3.1.21.10 crossover junction endodeoxyribonucleaseSpecify your search results
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- 3.1.21.10
-
fork
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helicase
- brca2
-
stall
-
nucleases
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rad51d
-
endonucleases
-
structure-specific
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germline
-
duplex
-
four-way
-
meiotic
-
nick
-
supercoiled
- reca
-
chromatid
-
transposition
- transposase
-
fanconi
-
restart
-
recombinases
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interstrand
-
cointegrate
- bard1
-
heteroduplexes
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structure-selective
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topoisomerases
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nucleolytic
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recbcd
- mre11
-
mutyh
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d-loops
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break-induced
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gammadelta
-
catenane
-
religation
-
post-replicative
-
recombination-dependent
-
fancm
-
four-armed
-
salpingo-oophorectomy
- medicine
The enzyme appears in viruses and cellular organisms
Reaction Schemes
endonucleolytic cleavage at a junction such as a reciprocal single-stranded crossover between two homologous DNA duplexes (Holliday junction)
Synonyms
rad51c, resolvase, mus81, mus81-eme1, mus81-mms4, ruvabc, t7 endonuclease i, endonuclease i, holliday junction resolvase, endonuclease vii, 
more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
BpuJI
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BpuJI is cleaved into a N-terminal domain, NTD, binding domain, and a C-terminal domain, CTD, catalytic domain, CTD is structurally related to archaeal Holliday junction resolvases
crossover junction endodeoxyribonuclease
cruciform-cutting endonuclease
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DNA HJ resolvase
EC 3.1.22.4
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formerly
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Eme1
endonuclease RuvC
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endonuclease VII
endonuclease X3
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GEN1
HJ resolvase
HJ resolving enzyme
HJ-resolving nuclease
Hjc
Hjc protein
Hje
Hjr
Holliday juction resolvase ruvC
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Holliday junction endonuclease CCE1
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Holliday junction nuclease ruvC
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Holliday junction resolvase
Holliday junction resolvase Yen1
Holliday junction resolving enzyme
Holliday junction resolving enzyme Hjc
Holliday junction resolving enzyme Hje
Holliday junction-cleaving endonuclease
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Holliday junction-resolving endoribonuclease
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Holliday junction-resolving enzyme
Holliday junction-resolving enzymes
junction-resolving enzyme
MBP-RuvC
MOC1
MUS-81
Mus81
Mus81 endonuclease
Mus81 nuclease
Mus81-Eme1
Mus81-Eme1/Mms4
MUS81-EME1A complex
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cleaves 3'-flap structures, nicked Holliday junctions, and, with reduced efficiency, intact Holliday junctions
MUS81-EME1B complex
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cleaves 3'-flap structures, nicked Holliday junctions, and, with reduced efficiency, intact Holliday junctions
phage T7 endonuclease I
phage T7 junction-resolving enzyme endonuclease I
RAD51C
RecU
RecU HJ resolvase
RecU Holliday junction resolvase
RecU Holliday-junction resolvase
RecU protein
RecUMge
resolving enzyme CCE1
RusA endonuclease
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RusA Holliday junction resolvase
RuvA
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RuvA is a Holliday junction-specific DNA-binding protein and facilitates the interaction of RuvB with the junction
RuvAB complex
ruvABC
RuvABC complex
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resolves Holliday junctions to produce recombinant molecules
RuvC
RuvC endonuclease
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RuvX
SLX-1
Slx1
Slx4
additional information
crossover junction endodeoxyribonuclease
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crossover junction endodeoxyribonuclease
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Holliday junction resolvase
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Holliday junction resolvase
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Mus81 endonuclease
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Mus81 is the catalytic component of the heterodimeric endonuclease Mus81-Eme1
resolving enzyme CCE1
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RusA Holliday junction resolvase
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RusA Holliday junction resolvase
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the enzyme is encoded by a cryptic rusA gene of the defective pyrophage DLP12
RuvX
Holliday junction resolvase formed by dimerisation of the monomeric YqgF nuclease domain
additional information
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the enzyme belongs to the nuclease superfamily, and is clearly related to the restriction enzymes
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
endonucleolytic cleavage at a junction such as a reciprocal single-stranded crossover between two homologous DNA duplexes (Holliday junction)
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric ester
hydrolysis of phosphoric ester
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CAS REGISTRY NUMBER
COMMENTARY
99676-43-4
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
5'-flapped junction in DNA + H2O
?
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the enzyme shows a robust activity with a specific cleavage site in the 5'-overhang strand exactly one nucleotide 3' of the branch point
-
-
?
DNA junction 1 + H2O
?
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DNA junction 1 substrate assembled from strands b50, h50,r50,x50, r55, b1-27, b28-50
-
-
?
restriction fork in DANN + H2O
?
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GEN1 preferentially cleaves strand 1 exactly at the branch point, whereas SEND1 preferentially cleaves one nucleotide in the 3' direction of the branch point. Cleavage in strand 2 is not detected with either GEN1 or SEND1 indicating a preference for cleavage of the lagging strand matrix
-
-
?
synthetic DNA junction
?
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cleavage of bimobile Holliday junctions occurs by introduction of nicks symmetrically at the 3'-side of thymine. The enzyme recognizes mainly topological symmetry of the Holliday junction but not the sequence symmetry per se so that a long homologous core sequence is not essential for cleavage
-
-
?
synthetic Holliday junction X10
?
synthetic DNA junction, containing the original 12 base pair core of homology, in which the region of homology is extended to a length of 10 base pairs
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-
?
synthetic Holliday junction X12
?
synthetic DNA junction, containing the original 12 base pair core of homology, in which the region of homology is extended to a length of 12 base pairs
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?
synthetic Holliday junction X26
?
synthetic DNA junction, containing the original 12 base pair core of homology, in which the region of homology is extended to a length of 26 base pairs
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?
DNA + H2O
?
cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. CFK1a01 is cleaved along both axes with equal efficiency, cleavage positions
-
?
DNA + H2O
?
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cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. For the substrate CFK1a01 the enzyme has a strong bias for the 01/03 axis, cleavage positions
-
?
DNA + H2O
?
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cleavage of Holliday junctions, different DNA substrates, 4-strand, 3-strand, Y-junction, flayed structure, 50-bp, 50-nt, 194-bp and 194-nt, annealing of ssDNA
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-
?
DNA + H2O
?
Holliday structure, RecU acts by making two simultaneous cuts in the phosphodiester backbone via its two active sites
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?
DNA + H2O
?
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Holliday structure, RecU stimulates RecA binding to ssDNA and RecA-catalyzed D-loop formation but inhibits RecA-mediated three-strand exchange reaction and ssDNA-dependent dATP or rATP hydrolysis
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-
?
DNA + H2O
?
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Holliday structure, RecU stimulates RecA binding to ssDNA and RecA-catalyzed D-loop formation but inhibits RecA-mediated three-strand exchange reaction and ssDNA-dependent dATP or rATP hydrolysis
-
-
?
DNA + H2O
?
-
32P-labeled four-way DNA junction J1 or junction Z1
-
?
DNA + H2O
?
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RuvC binding induces a twofold symmetrical X-shaped Holliday junction structure with two alternative conformers. The enzyme uses multivalency to keep the Holliday junction dynamic
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-
?
DNA + H2O
?
the enzyme uses multivalency to keep the Holliday junction dynamic
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?
DNA + H2O
?
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Holliday structure, RAD51C involved in HJ processing, catalysis of HJ resolution and ATP-dependent branch migration
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?
DNA + H2O
?
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intriguing substrate preference for nicked Holliday junctions and D-loops, generating crossovers
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?
DNA + H2O
?
GEN1 contains a chromodomain as an additional DNA interaction site. The GEN1 chromodomain directly contacts DNA and its truncation severely hampers GEN1's catalytic activity
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-
?
DNA + H2O
?
the enzyme uses multivalency to keep the Holliday junction dynamic
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?
DNA + H2O
?
Lambdavirus lambda
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Rap mediates symmetrical resolution of 50bp and chi Holliday structures containing larger homologous cores
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?
DNA + H2O
?
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cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. CFK1a01 is cleaved more efficiently along the 01/04 axis, cleavage positions
-
?
DNA + H2O
?
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cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. CFK1a01 is cleaved along both axes with equal efficiency, cleavage positions
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?
DNA + H2O
?
cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. CFK1a01 is cleaved along both axes with equal efficiency, cleavage positions
-
?
DNA + H2O
?
a fixed junction with heterologous arms and a defined point of strand exchange. Holliday junction resolving enzyme Hjc cleaves all four strands of the junction, three nucleotides 3' of the point of the strand exchange. Holliday junction resolving enzyme Hjr cleaves all four arms of the junction. Significant cleavage occurs one nucleotide 3' of the point of strand exchange
-
?
DNA + H2O
?
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the enzyme efficiently cleaves four-way junctions and less efficiently three-way junctions. The cleaved strand is rejoined by ligation, which means that cleavage occurs at the symmetrically related sites of the two strands, to leave 5'-phosphate and 3'-hydroxyl termini
-
?
DNA + H2O
?
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cleaves the Holliday junction in a symmetrical mode by introducing two nicks in opposite strands across the junction. Cleavage of fixed cruciform DNA CFK1a11 and CFK1a01 which allows a branch migration over ten nucleotides. The pair of opposite strands are 01/0.3 and 02/04 in CFK1a01 and 11/13 and 12/14 in CFK1a11. CFK1a11 is cleaved at identical positions, three nucleotides 3' of the junction. Strong bias for cleavage axis 12/14 with more than 80% of the substrate being cleaved, while only less than 10% along axis 11713 is cleaved. CFK1a01 is cleaved along both axes with equal efficiency, cleavage positions
-
?
DNA + H2O
?
a fixed junction with heterologous arms and a defined point of strand exchange. Holliday junction resolving enzyme Hjccleaves all four strands of the junction, three nucleotides 3' of the point of the strand exchange. Holliday junction resolving enzyme Hje cleaves only two strands, two nucleotides 3' of the junction centre. Hje is specific for strands that adopt a continous conformation in the stacked form of the junction
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?
DNA + H2O
?
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Holliday structure, junction Jbm5, catalytic site of Hje close to the N-terminus of strand betaB and a bend in betaC
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?
DNA + H2O
?
a fixed junction with heterologous arms and a defined point of strand exchange. Holliday junction resolving enzyme Hjccleaves all four strands of the junction, three nucleotides 3' of the point of the strand exchange. Holliday junction resolving enzyme Hje cleaves only two strands, two nucleotides 3' of the junction centre. Hje is specific for strands that adopt a continous conformation in the stacked form of the junction
-
?
DNA + H2O
?
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intriguing substrate preference for nicked Holliday junctions and D-loops, generating crossovers
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-
?
DNA + H2O
?
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resolves the synthetic four-way junction X12
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?
DNA + H2O
?
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the enzyme is required for mtDNA transmission and affects mtDNA content
-
?
DNA + H2O
?
Tequatrovirus T4
cruciform DNA substrate CF110. High affinity for the Holliday structure containing one nick
-
?
DNA + H2O
?
Thermochaetoides thermophila
binding to junctions in dimeric form, introducing symmetrical bilateral cleavages, the second of which is accelerated to promote productive resolution
-
-
?
DNA + H2O
?
Thermochaetoides thermophila
the enzyme uses multivalency to keep the Holliday junction dynamic
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-
?
DNA + H2O
?
Thermochaetoides thermophila DSM 1495
binding to junctions in dimeric form, introducing symmetrical bilateral cleavages, the second of which is accelerated to promote productive resolution
-
-
?
DNA + H2O
?
Thermochaetoides thermophila DSM 1495
the enzyme uses multivalency to keep the Holliday junction dynamic
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-
?
DNA + H2O
hydrolyzed DNA
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the enzyme from gene G44P preferentially cleaves Holliday junctions, but also, with lower efficiency, replicated D-loops
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-
?
DNA + H2O
hydrolyzed DNA
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at low concentrations the enzyme binds preferentially to the junction, in high concentrations it binds nonspecifically to any part of the DNA
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?
DNA + H2O
hydrolyzed DNA
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DNA topology rather than a sequence determine the cleavage site
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?
DNA + H2O
hydrolyzed DNA
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nicks the ssDNA strands across the junction at symmetrical positions within the homologous arms
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?
DNA + H2O
hydrolyzed DNA
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nicks the ssDNA strands across the junction at symmetrical positions within the homologous arms
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?
DNA + H2O
hydrolyzed DNA
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nicks the ssDNA strands across the junction at symmetrical positions within the homologous arms
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?
DNA + H2O
hydrolyzed DNA
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sequence specific: 5'- ATT - cleavage site - G-3', substrate: homologous core of 12 bp between the arms is required for cleavage but not for binding of the enzyme to the substrate, cleaves also Y-junctions with homologous core
-
?
DNA + H2O
hydrolyzed DNA
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sequence specificity: cleavage 5' of a CC-dinucleotide, binds weakly to linear duplex DNA
-
?
DNA + H2O
hydrolyzed DNA
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optimal sequence for cleavage: A equally well as T - TT - cutting site- C better than G or A, fastest when cleavage occurs at point of strand exchange, cleavage still possible 1 nucleotide 3' of this position when directed by the sequence, Y-junctions containing the sequence are cleavable
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?
DNA + H2O
hydrolyzed DNA
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cleavage at or near the point of strand exchange
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?
DNA + H2O
hydrolyzed DNA
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digestion of Bowtie junctions, Holliday junctions analogue containing 5',5' and 3',3' linkages in its crossover strands. The enzyme cleaves antiparralel junctions much more efficiently than parallel junctions where the protein can bind only one site at a time. The presence of two binding sites leads to communication between the two subunits of the enzyme to increase its activity
-
?
DNA + H2O
hydrolyzed DNA
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in contrast to GEN1, MUS81-EME1 cleaves intact Holliday junctions poorly (preferring nicked Holliday junctions, 3'-flaps, and replication fork structures as its DNA substrates). SLX1-SLX4 and MUS81-EME1 cooperatively cleave Holliday junctions by a nick and counter-nick mechanism
-
-
?
DNA + H2O
hydrolyzed DNA
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the enzyme cleaves a variety of DNA structures including intact Holliday junction and nicked and gapped duplex DNAs generating double-strand breaks. MUS81-EME2 cleaves two strands among three strands present in an nicked duplex. MUS81-EME2 preferentially cleaves nicked duplexes lacking a 5'-phosphate at the nick
-
-
?
DNA + H2O
hydrolyzed DNA
-
the substrate spectrum of MUS81-EME1 comprises 3'-flaps (duplex DNA with a 3'-single-stranded flap), double-stranded three-way junctions that resemble replication forks, Holliday junction precursors, and fully ligated Holliday junctions. Slx1Slx4 cleaves splayed arm DNA substrates (a duplex with unpaired 3'- and 5'-overhangs on one side), 5'-flaps (duplex DNA with a 5'-single-stranded flap), replication forks, and Holliday junctionss
-
-
?
DNA + H2O
hydrolyzed DNA
synthetic mobile four way junction Jbm5 as a substrate. The Hjc activity cleaves all four strands of the junction, three nucleotides 3' of the point of strand exchange. Pyrococcus has two Holliday junction resolving enzymes with different specificity: the cellular Hjc enzyme, and in addition the Hjr enzyme (probably of viral origin)
-
-
?
DNA + H2O
hydrolyzed DNA
synthetic mobile four way junction Jbm5 as a substrate. The Hjr enzyme cleaves all four arms of the junction significantly. Cleavage occurs one nucleotide 3' of the point of strand exchange. The organisms has two Holliday junction resolving enzymes with different specificity: the cellular Hjc enzyme, and in addition the Hjr enzyme (probably of viral origin)
-
-
?
DNA + H2O
hydrolyzed DNA
-
introduces paired nicks into opposing continous strands of a stacked X-junction, independent of local sequence, cleaves an Y-junction in the presence of a bulge in one strand so that stacking of the junction becomes possible
-
?
DNA + H2O
hydrolyzed DNA
synthetic mobile four way junction Jbm5 as a substrate. The Hjc activity cleaves all four strands of the junction, three nucleotides 3' of the point of strand exchange. Sulfolobus solfataricus has two Holliday junction resolving enzymes with different specificity: the cellular Hjc enzyme, and in addition the Hjr enzyme (probably of viral origin)
-
-
?
DNA + H2O
hydrolyzed DNA
synthetic mobile four way junction Jbm5 as a substrate. The Hjc activity cleaves all four strands of the junction, three nucleotides 3' of the point of strand exchange. Sulfolobus solfataricus has two Holliday junction resolving enzymes with different specificity: the cellular Hjc enzyme, and in addition the Hjr enzyme (probably of viral origin)
-
-
?
DNA + H2O
hydrolyzed DNA
-
nicks the ssDNA strands across the junction at symmetrical positions within the homologous arms
-
?
DNA + H2O
hydrolyzed DNA
-
Endo X3 cleaves Y-junctions,heteroduplex-loop DNA and VFS-DNA
-
?
DNA + H2O
hydrolyzed DNA
-
prefers cruciform base of X-junctions, Y-junctions are cleaved with a 5fold reduced efficiency, symmetric 6-bp sequence is needed
-
?
DNA + H2O
hydrolyzed DNA
-
specifically cleaves Holliday junctions, e.g. in bacteriophage G4 figure-8 molecules, cleavage at either of two sites present in the stem of the cruciform, not at the end of the stem
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme makes sequential cleavages in DNA junctions within the lifetime of the complex. The cleavage at a given site is accelerated by a factor of 5-10 when it occurs subsequently to the initial cleavage, thereby facilitating productive paired resolution cleavages
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme requires inherent rotational flexibility in DNA junctions for optimal catalysis. Recognition of 3'-flap and nicked Holliday junction substrates involves induction of a sharp bend with a 100° angle between two duplex DNA arms
-
-
?
DNA + H2O
hydrolyzed DNA
-
nicks favorably the continous strand at the point of strand exchange
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?
DNA + H2O
hydrolyzed DNA
-
binds to the X-junction in two specific complexes I and II, unfolds the stacked X-structure, cleaves at a subset of 5'-CT and 5'-TT sequences
-
?
DNA + H2O
hydrolyzed DNA
-
under physiological levels of salt it favors X-junctions, cleaves in vitro a range of other DNAs as Y-junction, flap, flayed, nicked, partial duplex, cleavage site always 3' of thymine nucleotides, at or one nucleotide 3' from the point of strand exchange
-
?
DNA + H2O
hydrolyzed DNA
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cleaves at the recognition sequence 5'-CT, preference for the point of strand exchange of fixed junctions
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme is required for meiotic crossing over but not for gene conversion
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme prevents mitochondrial DNA aggregation in Schizosaccharomyces pombe
-
?
DNA + H2O
hydrolyzed DNA
Tequatrovirus T4
-
DNA topology rather than a sequence determine the cleavage site
-
?
DNA + H2O
hydrolyzed DNA
Tequatrovirus T4
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performs 2 separate strand cleavages on the junction, cleaves a number of other structures that have in common bent helices
-
?
Holliday junction in DNA + H2O
?
-
both recombinant AtMUS81-EME1A and AtMUS81-EME1B complexes can cleave the intact Holliday junction, however, the nicked form serves as a better substrate for both of the homologous complexes and was cut with higher efficiency than the intact form
-
-
?
Holliday junction in DNA + H2O
?
the enzyme binds and cleaves the core of Holliday junctions symmetrically
-
-
?
Holliday junction in DNA + H2O
?
-
GEN1 resolves Holliday junctions by the introduction of symmetrically related cuts across the junction point, to produce nicked duplex products in which the nicks can be readily ligated, GEN1 leaves ligatable nicks after symmetrical cleavage
-
-
?
Holliday junction in DNA + H2O
?
-
RuvA protein binds Holliday junctions in preference to any other substrate
-
-
?
Holliday junction in DNA + H2O
?
Sulfolobus has two distinct junction resolving enzymes, Hjc and Hje, with differing substrate specificities. The Hje and Hjc activities display pronounced differences in their patterns of cleavage of the junction, and no nucleotide positions are cleaved by both enzymes. Whilst Hje introduces multiple nicks in each arm of the mobile junction Jbm5, Hjc cuts each arm at only a single site. Hje introduces paired nicks on only the continuous h and x strands of the fixed junction J3, two nucleotides 3' of the point of strand exchange. In contrast, recombinant Hjc cleaves all four strands of the junction, with cleavage three nucleotides 3' of the junction centre
-
-
?
Holliday junction in DNA + H2O
?
Sulfolobus has two distinct junction resolving enzymes, Hjc and Hje, with differing substrate specificities. The Hje and Hjc activities display pronounced differences in their patterns of cleavage of the junction, and no nucleotide positions are cleaved by both enzymes. Whilst Hje introduces multiple nicks in each arm of the mobile junction Jbm5, Hjc cuts each arm at only a single site. Hje introduces paired nicks on only the continuous h and x strands of the fixed junction J3, two nucleotides 3' of the point of strand exchange. In contrast, recombinant Hjc cleaves all four strands of the junction, with cleavage three nucleotides 3' of the junction centre
-
-
?
Holliday junction in DNA + H2O
?
-
Yen1 resolves Holliday junctions by the introduction of symmetrically related cuts across the junction point, to produce nicked duplex products in which the nicks can be readily ligated, Yen1 leaves ligatable nicks after symmetrical cleavage
-
-
?
Holliday junction X0 + H2O
?
-
GEN1 cuts the immobile junction X0 at a unique site located one nucleotide to the 3' side of the junction point
-
-
?
Holliday junction X0 + H2O
?
-
Yen1 cuts the immobile junction X0 at a unique site located one nucleotide to the 3' side of the junction point
-
-
?
additional information
?
-
-
a 3'-flap is not a suitable substrate
-
-
?
additional information
?
-
RecU interacts with RecA and inhibits its single-stranded DNA-dependent dATP hydrolysis
-
-
?
additional information
?
-
-
RecU interacts with RecA and inhibits its single-stranded DNA-dependent dATP hydrolysis
-
-
?
additional information
?
-
-
RecU has two activities: in concert with RuvAB, it catalyzes the resolution of Holliday junctions, and, alone, it modulates RecA activities. RecU does not modulate RecA when it is bound to the Holliday junction
-
-
?
additional information
?
-
-
ResT can use asymmetrized substrates that mimic the properties of a recombination site for a tyrosine recombinase, to form Holliday junctions
-
-
?
additional information
?
-
-
RuvA also stimulates RuvB helicase activity
-
-
?
additional information
?
-
-
RusA cleaves Holliday junctions with high specificity and efficiency, and has comparatively little activity on other forms of branched DNAs unless these can adopt a four-way branched configuration mimicking a Holliday junction
-
-
?
additional information
?
-
-
endonuclease I catalyses the breakage of the P-O3' bond
-
-
?
additional information
?
-
-
oriented cleavage on Holliday junction 1 by FPV resolvase. Active site residues are D7, E60, K102, D132, and D135. For the wild-type complex in the presence of EDTA or Ca2+, migration is consistent with the DNA arms arranged in near-tetrahedral geometry. However, the D7N active-site mutant resolvase holds the arms in a more planar arrangement in EDTA, Ca2+, or Mg2+ conditions, implicating metal-dependent contacts at the active site in the larger architecture of the complex
-
-
?
additional information
?
-
-
X12, nX12, 3'flap - EcME is able to convert all three substrates to linear duplex product
-
-
?
additional information
?
-
-
TRF2 contributes to t-loop stabilisation by stimulating Holliday junction formation and by preventing resolvase cleavage,TRF2 greatly increases the rate of Holliday junction formation and blocks the cleavage by various types of Holliday junction resolving activities, including the human GEN1 protein, the Myb-like domain of TRF2 slows the rate of junction migration, while the basic domain accelerates it
-
-
?
additional information
?
-
-
cleavage of a four-way DNA junction by the human SLX1-SLX4 complex, overview
-
-
?
additional information
?
-
-
substrate specificity of GEN1, and GEN1-Holliday junction complex structures, overview
-
-
?
additional information
?
-
-
the enzyme exhibits relaxed substrate specificity, cleaving a variety of branched DNA/RNA substrates. Notably, ATP hydrolysis plays a regulatory role, rendering the enzyme from a canonical Holliday junction resolvase to a DNA/RNA non-sequence specific endonuclease. The enzyme also has ATPase activity
-
-
-
additional information
?
-
-
the enzyme exhibits relaxed substrate specificity, cleaving a variety of branched DNA/RNA substrates. Notably, ATP hydrolysis plays a regulatory role, rendering the enzyme from a canonical Holliday junction resolvase to a DNA/RNA non-sequence specific endonuclease. The enzyme also has ATPase activity
-
-
-
additional information
?
-
-
RecU binds in a nonspecific fashion to Holliday junction substrates and, in the presence of Mn2+, cleaves these substrates at a specific sequence 5'-G/TC2C/TTA/GG-3'. Amino acid residues E11, K31, D57, Y58, Y66, D68, E70, K72, T74, K76, Q88, and L92 may play either a direct or indirect role in the catalysis of Holliday junction resolution
-
-
?
additional information
?
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
-
RecU binds in a nonspecific fashion to Holliday junction substrates and, in the presence of Mn2+, cleaves these substrates at a specific sequence 5'-G/TC2C/TTA/GG-3'. Amino acid residues E11, K31, D57, Y58, Y66, D68, E70, K72, T74, K76, Q88, and L92 may play either a direct or indirect role in the catalysis of Holliday junction resolution
-
-
?
additional information
?
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
-
the enzyme exhibits cytosine-dependent Holliday junction resolution of a synthetic bimobile Holliday junction of a 2-base-pair homologous core with a CCGG core. The enzyme has only weak cleavage activity against synthetic bimobile Holliday junctions of a 2-base-pair homologous core with CGCG and CATG cores
-
-
-
additional information
?
-
-
the enzyme specifically binds to the Holliday junction DNA and preferentially cleaves it at the consensus 5'-TTC-3'
-
-
-
additional information
?
-
-
class I substrates reflect low Km and high kcat and include the nicked Holliday junction, 3'-flapped and replication fork-like structures
-
-
?
additional information
?
-
-
class II substrates share low Km but low kcat relative to class I substrates and include the D-loop and partial Holliday junction
-
-
?
additional information
?
-
-
class III substrates are defined by splayed Y junctions having high Km and low Kcat
-
-
?
additional information
?
-
ATPase SisPINA interacts with SisHjc and coordinates Holliday junction migration and cleavage
-
-
?
additional information
?
-
ATPase SisPINA interacts with SisHjc and coordinates Holliday junction migration and cleavage
-
-
?
additional information
?
-
-
enzyme substrate specificity, overview. A specific interaction occurs between SIRV2 Holliday junction resolving enzyme, Hjr, and the SIRV2 virion body coat protein, SIRV2gp26
-
-
?
additional information
?
-
-
usage of two plasmids containing hairpin four-way junctions constructed to assay resolving enzyme activity
-
-
?
additional information
?
-
Tequatrovirus T4
-
activity with different constructs of Y-DNA under various packaging conditions, T4 endo VII resolvase cleavage sites in Y-DNAs, overview
-
-
?
additional information
?
-
Thermochaetoides thermophila
the enzyme is highly selective for four-way DNA junctions, cleaving 1 nucleotide 3' to the point of strand exchange on two strands symmetrically disposed about a diagonal axis
-
-
-
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
DNA + H2O
?
-
the enzyme is required for mtDNA transmission and affects mtDNA content
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme from gene G44P preferentially cleaves Holliday junctions, but also, with lower efficiency, replicated D-loops
-
-
?
DNA + H2O
hydrolyzed DNA
-
in contrast to GEN1, MUS81-EME1 cleaves intact Holliday junctions poorly (preferring nicked Holliday junctions, 3'-flaps, and replication fork structures as its DNA substrates). SLX1-SLX4 and MUS81-EME1 cooperatively cleave Holliday junctions by a nick and counter-nick mechanism
-
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme cleaves a variety of DNA structures including intact Holliday junction and nicked and gapped duplex DNAs generating double-strand breaks. MUS81-EME2 cleaves two strands among three strands present in an nicked duplex. MUS81-EME2 preferentially cleaves nicked duplexes lacking a 5'-phosphate at the nick
-
-
?
DNA + H2O
hydrolyzed DNA
-
the substrate spectrum of MUS81-EME1 comprises 3'-flaps (duplex DNA with a 3'-single-stranded flap), double-stranded three-way junctions that resemble replication forks, Holliday junction precursors, and fully ligated Holliday junctions. Slx1Slx4 cleaves splayed arm DNA substrates (a duplex with unpaired 3'- and 5'-overhangs on one side), 5'-flaps (duplex DNA with a 5'-single-stranded flap), replication forks, and Holliday junctionss
-
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme requires inherent rotational flexibility in DNA junctions for optimal catalysis. Recognition of 3'-flap and nicked Holliday junction substrates involves induction of a sharp bend with a 100° angle between two duplex DNA arms
-
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme is required for meiotic crossing over but not for gene conversion
-
?
DNA + H2O
hydrolyzed DNA
-
the enzyme prevents mitochondrial DNA aggregation in Schizosaccharomyces pombe
-
?
additional information
?
-
-
RecU has two activities: in concert with RuvAB, it catalyzes the resolution of Holliday junctions, and, alone, it modulates RecA activities. RecU does not modulate RecA when it is bound to the Holliday junction
-
-
?
additional information
?
-
-
RuvA also stimulates RuvB helicase activity
-
-
?
additional information
?
-
-
RusA cleaves Holliday junctions with high specificity and efficiency, and has comparatively little activity on other forms of branched DNAs unless these can adopt a four-way branched configuration mimicking a Holliday junction
-
-
?
additional information
?
-
-
endonuclease I catalyses the breakage of the P-O3' bond
-
-
?
additional information
?
-
-
cleavage of a four-way DNA junction by the human SLX1-SLX4 complex, overview
-
-
?
additional information
?
-
-
substrate specificity of GEN1, and GEN1-Holliday junction complex structures, overview
-
-
?
additional information
?
-
-
the enzyme exhibits relaxed substrate specificity, cleaving a variety of branched DNA/RNA substrates. Notably, ATP hydrolysis plays a regulatory role, rendering the enzyme from a canonical Holliday junction resolvase to a DNA/RNA non-sequence specific endonuclease. The enzyme also has ATPase activity
-
-
-
additional information
?
-
-
the enzyme exhibits relaxed substrate specificity, cleaving a variety of branched DNA/RNA substrates. Notably, ATP hydrolysis plays a regulatory role, rendering the enzyme from a canonical Holliday junction resolvase to a DNA/RNA non-sequence specific endonuclease. The enzyme also has ATPase activity
-
-
-
additional information
?
-
-
RecU binds in a nonspecific fashion to Holliday junction substrates and, in the presence of Mn2+, cleaves these substrates at a specific sequence 5'-G/TC2C/TTA/GG-3'. Amino acid residues E11, K31, D57, Y58, Y66, D68, E70, K72, T74, K76, Q88, and L92 may play either a direct or indirect role in the catalysis of Holliday junction resolution
-
-
?
additional information
?
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
-
RecU binds in a nonspecific fashion to Holliday junction substrates and, in the presence of Mn2+, cleaves these substrates at a specific sequence 5'-G/TC2C/TTA/GG-3'. Amino acid residues E11, K31, D57, Y58, Y66, D68, E70, K72, T74, K76, Q88, and L92 may play either a direct or indirect role in the catalysis of Holliday junction resolution
-
-
?
additional information
?
-
RecUMge binds Holliday junction substrates and large double-stranded DNA molecules and cleaves Holliday junction substrates at the sequence 5'-G/TC-/-C/TTA/GG-3' in the presence of Mn2+, cleavage site determination, overview. RecUMge does not bind to small, linear dsDNA substrates. RecUMge cleavage of alternatively branched substrates, overview
-
-
?
additional information
?
-
ATPase SisPINA interacts with SisHjc and coordinates Holliday junction migration and cleavage
-
-
?
additional information
?
-
ATPase SisPINA interacts with SisHjc and coordinates Holliday junction migration and cleavage
-
-
?
additional information
?
-
-
enzyme substrate specificity, overview. A specific interaction occurs between SIRV2 Holliday junction resolving enzyme, Hjr, and the SIRV2 virion body coat protein, SIRV2gp26
-
-
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Co2+
divalent cations
Mg2+
Mn2+
Zn2+
additional information
Ca2+
-
binding of two tetramers of RuvA on the junction is favored by divalent cations, Ca2+ enhances the binding of RuvA to the junction
Ca2+
-
cannot replace Mg2+ or Mn2+ at concentrations between 1-500 mM
Ca2+
-
Endo X3, 70% of the maximal activity obtained when Mg2+ is replaced by Ca2+
Co2+
-
Endo X3 shows less than 10% of full activity at a concentration of 10 mM Co2+
Mg2+
-
the endonuclease activity is optimal for the intact Holliday junction substrate at a range of 1 mM Mg2+ for both of the homologous complexes, a decrease in activity is observed at higher concentrations, Mg2+ can be replaced by Ca2+
Mg2+
-
modulates affinity of RecU for DNA, at low concetrations RecU binds to 3- and 4-strands with 12- and 46-fold higher affinity than to 194-nt or 194-bp DNA, and with > 50-fold higher affinity than to flayed, Y-junction, 50-nt and 50-bp DNA substrates, at high concentrations, RecU introduces specific cuts on mobile 4-strand substrates
Mg2+
the enzyme requires high concentrations of Mg2+ (10-15 mM) for activity
Mg2+
-
optimum: 8-25 mM, binding per se is independent of Mg2+, cleavage mechanism depends on Mg2+
Mg2+
-
the enzyme binds to stacked-X junctions in the presence of cations, in the absence of cations the enzyme binds to square-planar junctions
Mg2+
the enzyme cleaves surface-immobilized Holliday junctions in presence of Mg2+ but not in Ca2+. In the presence of Mg2+, Holliday junctions fold into two alternatively stacked conformers. A single Holliday junction can undergo conformer exchange between U1 and U2 and the exchange rate decreases with increasing Mg2+ concentration
Mg2+
-
a Mg2+ ion is present in each of the two active sites in the homodimeric enzyme
Mg2+
Lambdavirus lambda
-
required for chi DNA cleavage, 0.1 mM is sufficient to promote stacking of the junction arms, at 1 mM no Rap-junction complexes were detected
Mg2+
the enzyme requires a divalent cation for Holliday junction cleavage (typically Mg2+ or Mn2+), but not for Holliday junction binding activity
Mg2+
-
optimum: 15-30 mM MgCl2, 10% of maximal activity in 4 mM MgCl2
Mg2+
strongly dependent on the presence of magnesium, with 15 mM Mg2+ ions optimal. Strong influence of magnesium ion concentrations on protein-junction complex stability. A dramatic shift in binding affinity is observed, with apparent dissociation constants for binding in the presence of 1.5, 2.5, 5 and 15 mM magnesium ions of 76 nM, 530 nM, 1100 nM and 1700 nM, respectively
Mg2+
-
Endo X3, optimum: 10 mM MgCl2, in presence of 10 mM MgCl2 the enzyme is salt sensitive
Mn2+
-
supports activity as a cofactor to a higher extent then Mg2+ and leads to the formation of further cleavage products of lower molecular mass
Mn2+
Mn2+ substitutes as an efficient cofactor for cleavage by the enzyme, but does not stimulate resolution activity
Mn2+
-
preferred meta ion. The enzyme requires Mn2+ or Mg2+ as metal cofactors. Concentrations of Mn2+ above 3.3 mM inhibit cleavage
Mn2+
Lambdavirus lambda
-
required for chi DNA cleavage, reduced preference for cleavage in Mn2+ relative to Mg2+ on chi compared to small DNA substrates
Mn2+
the enzyme requires a divalent cation for Holliday junction cleavage (typically Mg2+ or Mn2+), but not for Holliday junction binding activity
Mn2+
-
replacement of MgCl2 by equimolar amounts of MnCl2 results in a 50% loss of activity
Mn2+
-
Endo X3, 70% of the maximal activity obtained when Mg2+ is replaced by Mn2+
Zn2+
-
cannot replace Mg2+ or Mn2+ at concentrations between 1-500 mM
additional information
-
Ni2+ and Zn2+ cannot serve as cofactors for catalytic activity
additional information
-
influence of metal cofactors on the activity and structure of the resolvase of fowlpox virus, overview. No activity with Ca2+, little activity with Cu2+, Fe2+, Ni2+, and Zn2+
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Cds1
-
the DNA replication checkpoint kinase Cds1 negatively regulates Mus81/Eme1 to preserve genomic integrity when replication is perturbed
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
PCNA protein
-
Holliday junction-resolving enzyme Hjc interacts physically with PCNA via a canonical C-terminal PIP motif. This interaction stimulates the junction cleavage activity of Hjc in vitro. PCNA is a toroidal protein that acts as a processivity factor for many proteins. In Sulfolobus solfataricus, PCNA is a heterotrimer that can function as a molecular toolbelt, forming simultaneous interactions with up to three partner proteins with related functions in a DNA-processing pathway
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00000052
intact Holliday junction
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
0.00000102
3'-flapped junction in DNA
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.037
intact Holliday junction
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
0.18
nX12 junction in single-stranded DNA
-
the nX12 substrate has four 25-bp duplex DNA arms, with a 12-bp homologous core that allows branch migration of the junction, and contains a nick at the cross-over point
-
0.0047
X12 junction in single-stranded DNA
-
like nX12, without the nick at the cross-over point
-
0.12
3'-flapped junction in DNA
-
the 3'flap substrate contains 50 bp of duplex DNA, a central nick, and 25 bases of single-stranded DNA
-
0.354
3'-flapped junction in DNA
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
0.0065
DNA junction 1
-
at 30°C, steady-state approximation under maximal velocity conditions at 15 mM MgCl2
-
0.05
DNA junction 1
-
at 65°C, pH 7, converting 6% of the substrate into product
-
0.000006
DNA junction 1 unconstrained
-
E226N, 480fold reduced turnover rate
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
346300
3'-flapped junction in DNA
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
71830
intact Holliday junction
-
in 25 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 0.2 mM dithiothreitol, at 37°C
-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
8
in the presence of Mn2+, resolution activity is optimal at physiological pH (about 8.0)
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
30
37
60
30
-
steady-state approximation at 0.30 min-1 under maximal velocity conditions at 15 mM MgCl2
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37 - 60
-
elevated temperatures (37-60°C) boost the catalytic activity of the enzyme
37 - 65
the optimum temperature for resolution is 65°C, with 70% of the substrate resolved within 15 min. At 37°C, less than 5% resolution is observed after 15 min, rising to a maximum level of resolution of 35% after 3 h
45 - 65
-
no multiple turnover catalysis at lower temperatures measurable due to the sensitivity of the assay
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Tequatrovirus T4
-
-
-
brenda
the enzyme RusA resolvase is encoded by a cryptic rusA gene of the defective pyrophage DLP12
-
-
brenda
-
664677, 666919, 683401, 683782, 684015, 692074, 694344, 714118, 715112, 729606, 729626, 730517, 730712, 731008
-
-
brenda
P9WGW3: RuvA, P9WGW1: RuvB, RuvAB Holliday junction resolvase complex
SwissProt
brenda
P9WGW3: RuvA, P9WGW1: RuvB, RuvAB Holliday junction resolvase complex
SwissProt
brenda
-
134317, 134323, 134327, 134330, 649970, 683336, 683401, 683942, 694125, 694344, 708314, 716393, 729606, 729626, 730518
-
-
brenda
-
-
-
brenda
Spcce1:ura4+ insertion mutant strain devoid of the Holliday junction resolvase activity
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
PSNG13, Bloom's syndrome-associated, BLM, helicase-deficient fibroblasts, PSNF5, BLM-complemented fibroblasts
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
wild type enzyme and truncated enzyme forms DELTA1-35 and DELTA1-35/CDELTA15
-
brenda
additional information
additional information
-
in exponentially growing cells RecU is present throughout the cells, after addition of 100ng/mol MMC discrete focus in the nucleoid, in 2.7% of the cells two foci
-
brenda
additional information
-
in exponentially growing cells RecU is present throughout the cells, after addition of 100ng/mol MMC discrete focus in the nucleoid, in 2.7% of the cells two foci
-
-
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
metabolism
physiological function
additional information
malfunction
-
yen1DELTA mutants are repair-proficient. Yen1DELTA mus81ELTA double mutant displays a more severe repair-defective phenotype than the mus81DELTA mutant. Yen1DELTA cells do not exhibit any obvious sensitivity to DNA-damaging agents like the wild-type, whereas yen1DELTA mus81DELTA double mutants are exquisitely sensitive to a variety of DNA-damaging agents that disturb replication fork progression. Yen1DELTA mus81DELTA cells show a hypersensitivity to all agents for which the mus81DELTA single mutant is sensitive: hydroxyurea, 4-nitroquinoline 1-oxide, phleomycin, camptothecin, UV-light, nitrogen mustard and cisplatin. Neither the yen1DELTA or mus81DELTA single mutants nor the yen1DELTA mus81DELTA double mutant show any sensitivity to ionizing radiation up to 200 Gy. Yen1DELTA sgs1DELTA cells are viable and exhibit a similar methyl methanesulfonate, hydroxyurea and 4-nitroquinoline 1-oxide-sensitivity to that observed with the sgs1DELTA single mutant. Toxic recombination intermediates accumulate in the absence of Yen1 and Mus81. After methyl methanesulfonate treatment, yen1DELTA mus81DELTA double mutants arrest with a G2 DNA content and unsegregated chromosomes. Overexpression of Yen1 partially rescues the methyl methanesulfonate sensitivity of mus81DELTA, Yen1 is the only member of the Rad2 family of nucleases that can complement mus81 defects. Yen1ELTA mus81ELTA double mutants are synthetically sick: 15-20 min increase in the duration of the cell cycle of yen1DELTA mus81DELTA double mutants compared with wild-type, yen1DELTA or mus81DELTA cells growing in YPD. Constitutive expression of Yen1-3xHA, but not the catalytically dead version of Yen1, reduces the doubling time of the yen1DELTA mus81DELTA mutant to wild-type levels
malfunction
-
gen-1 mutants are defective in DNA damage-dependent cell cycle arrest and apoptosis and in positional cloning of GEN-1, phenotypes, detailed overview
malfunction
-
single mutants of hjc and hef display no significant defects in growth or homologous recombination. Deletion of hef confers only moderate sensitivity to DNA crosslinking agents, whereas mutation of FANCM in leads to hypersensitivity in eukaryotes. Absence of hef or hjc leads to growth defects in presence of mitomycin C, deletion of both leads to synthetic lethality
malfunction
-
strains lacking RuvABC are inviable and accumulate Holliday junctions that interfere with growth and division of the cells
malfunction
-
yeast strains deleted for both YEN1 and MMS4 show a reduction in growth rate, and are very sensitive to DNA-damaging agents. Yeast cells are unable to carry out meiosis in the absence of both resolvases
malfunction
-
enzyme depletion results in the appearance of cells with compact nucleoids, septa formed over the DNA and anucleate cells. Enzyme-depleted cells also show increased septal recruitment of the DNA translocase SpoIIIE, presumably to resolve chromosome segregation defects. Additionally cells are more sensitive to DNA damaging agents such as mitomycin C or UV radiation
malfunction
-
GEN1 depletion results in aberrant centrosome numbers associated with the formation of multiple spindle poles in mitosis, an increased number of cells with multi-nuclei, increased apoptosis and an elevated level of spontaneous DNA damage
malfunction
-
loss of Mus81 activity sensitizes yeast cells to a large panel of agents whose effects inhibit RF progression, including methyl methanesulfonate, UV light, camptothecin, hydroxyurea, and DNA cross-linking agents
malfunction
-
the absence of SLX4 and BLM, or SLX4 and GEN1, is synthetically lethal in undamaged human cells
malfunction
mutations at the Cre04.g218000 gene are responsible for the mocH72 phenotype and with monokaryotic chloroplasts, which possess only a single chloroplast nucleoid and show unequal segregation of chloroplast nucleoids during chloroplast divisions
malfunction
reduced or no expression of MOC1 in Arabidopsis thaliana leads to growth defects and aberrant chloroplast nucleotide segregation
malfunction
-
enzyme depletion induces ectopic recombination between short dispersed repeats in plastidial DNA and disorganizes thylakoid membranes in plastid
malfunction
-
enzyme depletion induces ectopic recombination between short dispersed repeats in plastidial DNA and disorganizes thylakoid membranes in plastid
malfunction
-
enzyme depletion results in the appearance of cells with compact nucleoids, septa formed over the DNA and anucleate cells. Enzyme-depleted cells also show increased septal recruitment of the DNA translocase SpoIIIE, presumably to resolve chromosome segregation defects. Additionally cells are more sensitive to DNA damaging agents such as mitomycin C or UV radiation
-
metabolism
-
gen-1 acts in a non-canonical DNA damage checkpoint pathway
metabolism
-
Holliday junction resolving enzyme is a key enzyme in SIRV2 genome replication. Modeling linking SIRV2 Holliday junction resolving enzyme genome resolution to viral particle assembly, overview
physiological function
-
Yen1 and Mus81-Mms4 provide alternative and/or overlapping pathways for the repair of methyl methanesulfonate-induced DNA lesions. Yen1 can act upon recombination/repair intermediates that arise in MUS81-defective cells following replication fork damage
physiological function
-
four-way DNA Holliday junctions are resolved into duplex species by the action of the junction-resolving enzymes, nucleases selective for the structure of helical branchpoints. Mechanism of structural selectivity of these enzymes, overview
physiological function
-
four-way DNA Holliday junctions are resolved into duplex species by the action of the junction-resolving enzymes, nucleases selective for the structure of helical branchpoints. Mechanism of structural selectivity of these enzymes, overview
physiological function
-
Hef, a XPF/MUS81 family member found in Euryarchaea and is related to the Fanconi anemia protein FANCM, is essential for cell viability when the Holliday junction resolvase Hjc is absent, and both the helicase and nuclease activities of Hef are indispensable. Hef and Hjc provide alternative means to restart stalled DNA replication forks
physiological function
-
Holliday junction resolution is essential for chromosome segregation at meiosis and the repair of stalled/collapsed replication forks in mitotic cells. Resolution by introduction of symmetrically related nicks in two strands at, or close to, the junction point. Structural basis and mechanism of Holliday junction resolution by the human GEN1 protein, overview
physiological function
-
Holliday junctions need to be resolved to allow chromosome segregation, they are formed during homologous recombination. Both Yen1 and Mms4/Mus81 play important but not identical roles during vegetative growth and in meiosis. Yen1 and Mms4/Mus81 act as alternative resolvases for recombination events in meiosis, and the activity of at least one of them is essential to ensure proper meiosis and sporulation
physiological function
-
identification of a Caenorhabditis elegans dual function DNA double-strand break repair and DNA damage signaling protein orthologous to the human GEN1 Holliday junction resolving enzyme. GEN-1 is required for DNA double-strand break repair. The DNA damage-signaling function of GEN-1 is separable from its role in DNA repair. GEN-1 promotes germ cell cycle arrest and apoptosis via a pathway that acts in parallel to the canonical DNA damage response pathway mediated by RPA loading, CHK1 activation, and CEP-1/p53-mediated apoptosis induction. GEN-1 acts redundantly with the 9-1-1 complex to ensure genome stability. GEN-1 might act as a dual function Holliday junction resolvase that may coordinate DNA damage signaling with a late step in DNA double-strand break repair
physiological function
Tequatrovirus T4
-
in vivo the T4 phage packaging motor deals with Y- or X-structures in the replicative concatemer substrate by employing a portal-bound Holliday junction resolvase that trims and releases these DNA roadblocks to packaging. Purified T4 gp49 endonuclease VII resolvase can release DNA compression in vitro in prohead portal packaging motor anchored and arrested Y-DNA substrates. Conformational changes in both the motor proteins and the DNA substrate itself that are associated with the power stroke of the motor are consistent with a proposed linear motor employing a terminal-to-portal DNA grip-and-release mechanism
physiological function
-
molecular mechanism for the Holliday junction resolving enzyme four-way junction cleavage bias, minimal requirements for four-way junction cleavage, and substrate specificity. The cleavage is specific to four-way DNA junctions and inactive on other branched DNA molecules
physiological function
-
poxvirus DNA replication generates linear concatemers containing many copies of the viral genome with inverted repeat sequences at the junctions between monomers. The inverted repeats refold to generate Holliday junctions, which are cleaved by the virus-encoded resolvase enzyme to form unit-length genomes
physiological function
RecUMge is a Holliday junction resolvase that may play a central role in recombination in Mycoplasma genitalium
physiological function
-
RuvABC resolves Holliday junctions, with RuvAB driving branch migration and RuvC catalyzing junction cleavage
physiological function
-
the RecU protein has two activities: to recognize, distort, and cleave four-stranded recombination intermediates and to modulate RecA activities. The RuvB interaction and the catalytic residues are located in the cap region of dimeric RecU, while the stalk region is essential not only for RecA modulation but also for Holliday junction recognition
physiological function
-
the Sgs1-Top3-Rmi1 complex constitutes the main pathway for the processing of Holliday junction-containing homologous recombination repair intermediates but that Mus81-Mms4 can also resolve these intermediates. These intermediates are slowly resolved at the restrictive temperature, revealing a redundant resolution activity when Rmi1 is impaired. This resolution depends on Mus81-Mms4 but not on either Slx1-Slx4 or another HJ resolvase, Yen1
physiological function
the enzyme is involved in the pathway for homologous recombination of the cellular genome, which includes the RadA protein for strand exchange, and the Hjc protein for junction resolution
physiological function
the enzyme is involved in the pathway for homologous recombination of the cellular genome, which includesthe RadA protein for strand exchange, and the Hjc protein for junction resolution
physiological function
-
BLM, SLX4, MUS81, and GEN1 cooperate in replication fork maintenance. BTR, SLX-MUS, and GEN1 are important for chromosome stability and faithful chromosome segregation
physiological function
-
resolvase-mediated Holliday junction resolution safeguards chromosome segregation
physiological function
-
resolvase-mediated Holliday junction resolution safeguards chromosome segregation
physiological function
-
RuvC cleaves Holliday junctions by the introduction of two symmetrically related nicks. The enzyme also cuts branched DNA intermediates that originate directly from blocked replication forks, targeting them for origin-independent replication restart
physiological function
-
the enzyme functions in homology driven repair of DNA double strand breaks. Centrosome association but not catalytic activity of GEN1 is required for preventing centrosome hyper-amplification, formation of multiple mitotic spindles, and multi-nucleation
physiological function
-
the enzyme is required for chromosome segregation and DNA damage repair
physiological function
-
the enzyme plays a role in replication restart during the transition to concatemeric viral replication
physiological function
MOC1 may mediate chloroplast nucleoid segregation in green plants by resolving Holliday junctions
physiological function
replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1. Yen1 provides a downstream survival pathway, along which toxic DNA intermediates that arise when the Dna2 helicase activity fails to respond adequately to replication fork stalling are resolved
physiological function
RuvAB-dependent Holliday junction resolution appears to participate in the repair of the DNA fragmentation that initiates the protein synthesis- and ROS-dependent killing pathway
physiological function
the enzyme is essential for maintaining the morphology of chloroplast nucleotides
physiological function
-
the enzyme functions in the double-strand break repair and resolves Hollidays in mitochondria
physiological function
-
the enzyme functions in the double-strand break repair and resolves Hollidays in mitochondria
physiological function
-
the enzyme is involved in the pathway for homologous recombination of the cellular genome, which includes the RadA protein for strand exchange, and the Hjc protein for junction resolution
-
physiological function
-
replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1. Yen1 provides a downstream survival pathway, along which toxic DNA intermediates that arise when the Dna2 helicase activity fails to respond adequately to replication fork stalling are resolved
-
physiological function
-
the enzyme is required for chromosome segregation and DNA damage repair
-
physiological function
-
RuvAB-dependent Holliday junction resolution appears to participate in the repair of the DNA fragmentation that initiates the protein synthesis- and ROS-dependent killing pathway
-
physiological function
-
RecUMge is a Holliday junction resolvase that may play a central role in recombination in Mycoplasma genitalium
-
additional information
-
GEN1 is a monomeric 5'-flap endonuclease. The unique feature of GEN1 that distinguishes it from other Rad2/XPG nucleases is its ability to dimerize on Holliday junctions
additional information
Holliday junction resolution by RecUMge is Mn2+-, pH- and temperature-dependent
additional information
-
Holliday junction resolution by RecUMge is Mn2+-, pH- and temperature-dependent
additional information
-
recognition and manipulation of junction structure, overview
additional information
-
recognition and manipulation of junction structure, overview. Model of DNA bound in the active site of phage T7 endonuclease I
additional information
-
the RegG pathway is no alternative to the RuvABC pathway for resoling Holliday junctions, since the RecG pathway is very ineffective at removing junctions. A major function of RecG is to curb potentially pathological replication initiated via PriA protein at sites remote from oriC
additional information
-
Holliday junction resolution by RecUMge is Mn2+-, pH- and temperature-dependent
-
Show AA Sequence and Transmembrane Helices (44123 entries)
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
109000
-
homodimer, determined by sedimentation equilibrium experiments using an analytical ultracentrifuge
140000
-
determined by gel filtration using a 88 microM EcME solution, heterotetramer
40000
72000
73600
-
predicted molecular weight for His10-FLAG-Mus81, verified by PAGE
82000
-
determined by gel filtration using a 3 microM EcME solution, heterodimer
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
heterodimer
homodimer
monomer
monomer or dimer
tetramer
additional information
dimer
-
homodimer, 2 * 121930-135000, sedimentation and multi-angle light scattering
dimer
-
crystallographic 2fold rotation axis, Se-targeted SAD methods, dimeric in solution and in crystal
dimer
Thermochaetoides thermophila DSM 1495
-
-
-
dimer
-
homodimer, 67000-158000, gel filtration, chemical cross linking, sedimentation and multi-angle light scattering
homodimer
-
the endonuclease I forms an intimately associated symmetrical homodimer comprising two domains, each formed by residues 17-44 from one subunit and 50-145 from the other. The domains are connected by a bridge that forms part of an extended beta-sheet
monomer
-
GEN1 protein contains the XPG-N, XPG-I, and helix-hairpin-helix domains essential for nuclease activity, linked to a C-terminal tail region that appears to be naturally disordered
monomer or dimer
Thermochaetoides thermophila DSM 1495
-
-
-
additional information
-
the RecU structure shows a mushroom-like appearance, with a cap and a stalk region. The RuvB interaction and the catalytic residues are located in the cap region of dimeric RecU, while the stalk region is essential not only for RecA modulation but also for Holliday junction recognition
additional information
-
model of DNA bound in the active site of phage T7 endonuclease I
additional information
-
Mus81 and Eme1 are subunits of nuclear Holliday junction resolvase
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
additional information
phosphoprotein
-
the eukaryotic-like DNA repair proteins and protein kinases SiRe_0171, SiRe_2030, and SiRe_2056 are able to phosphorylate the enzyme in vitro. Enzyme phosphorylation by SiRe_0171 enhances the DNA repair capability
phosphoprotein
-
the eukaryotic-like DNA repair proteins and protein kinases SiRe_0171, SiRe_2030, and SiRe_2056 are able to phosphorylate the enzyme in vitro. Enzyme phosphorylation by SiRe_0171 enhances the DNA repair capability
-
additional information
-
cleavage activity of Mus81 is strictly regulated, part of this regulation occurs through post-translational modifications of Mus81
additional information
-
cleavage activity of Mus81 is strictly regulated, part of this regulation occurs through post-translational modifications of Mus81
additional information
-
cleavage activity of Mus81 is strictly regulated, part of this regulation occurs through post-translational modifications of Mus81
additional information
-
cleavage activity of Mus81 is strictly regulated, part of this regulation occurs through post-translational modifications of Mus81
additional information
-
cleavage activity of Mus81 is strictly regulated, part of this regulation occurs through post-translational modifications of Mus81
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
prismatic crystals obtained by sparse-matrix screening and sitting-drop vapour-diffusion, plate-like crystals by counter-diffusion in X-ray capillary
-
at 17°C, hanging drop method, in solution having a mushroom-like appearance, where the cap is formed by the main beta-sheet and flanking helices, structure of RecU superimposes best with PfHjc and archaeal HJR, the next best matches to RecU are a series of type II restriction endonucleases including EcoRV and Pvull
the structures of RusA-D70N and RusA-D70N-DNA complex are determined at resolutions of 1.2 A and 3.1 A, respectively
three distinct crystal forms, form I: triclinic space group P1, resolution: 4 A, form II: not grown to a size sufficient for X-ray diffraction, form III: monoclinic space group P2-1, resolution: 2.5 A
-
resolving the enzyme bound to DNA junctions, X-ray diffraction structure determination and analysis
-
improvements in crystallization conditions result in an increased resolution of the native enzyme crystal's diffraction data set of 1.4 A
-
the structure of Hjc is determined by using multiple-wavelength anomalous dispersion to a resolution of 2.15A
-
crystal structure of human HJ resolvase GEN1 complexed with DNA at 3.0 A resolution
resolving enzymes bound to DNA junctions, X-ray diffraction structure determination and analysis
-
apoenzyme, sitting drop vapor diffusion method, using 14% (w/v) PEG3350, 0.15 M NH4Cl. Mutant enzyme I112V/Q162K/E235Q/E239Q sitting drop vapor diffusion method, using 0.1M Bis-Tris, pH 6.3, 22% (w/v) PEG3350, 0.2 M Li2SO4. Mutant enzyme I112V/Q162K/E235Q/E239Q in complex with Holl8iday junctions, sitting drop vapor diffusion method, using 0.1 M sodium acetate trihydrate, pH 5.0, 8.5% (w/v) PEG3350, 2% Tacsimate, pH 4.0 or 0.1 M sodium acetate trihydrate, pH 5.6, 15% (w/v) 3-methyl-1,5-pentane diol, 2% (w/v) PEG4000, 2% (w/v) D-sorbitol
-
microbatch method with a silicone oil overlay, crystal structure at 2.0 A resolution
-
selenomethionine-containing enzyme crystallized by microbatch method with a silicone oil overlay, hexagonal form of the enzyme
-
crystal structure of Sulfolobus Hjc, determined by using multiple-wavelength anomalous dispersion, to a resolution of 2.15 A
-
sitting drop method, Mosquito nanolitre-drop crystallization robot, X-ray data collection
-
hanging drop vapor diffusion method, using 35% (w/v) polyethylene glycol 3350, 0.2M Li2SO4 and 0.1M Tris-HCl (pH 8.5)
in complex with a synthetic Holliday junction, using 0.1 M citric acid (pH 3.5) and 1.5 M NaCl in 0.3-0.5M ammonium phosphate
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
F81A
-
naturally occuring mutant, that is sensitive to DNA-damaging agents as a null recU strain, a severely impaired enzyme. The mutant poorly recognizes and distorts Holliday junctions. At high concentrations, RecUF81A binds to Holliday junctions but fails to cleave them. RecUF81A does not inhibit RecA dATPase and strand-exchange activities and it loses structural selectivity for X-shaped structures. Phenotype, detailed overview
K56A
mutant with a putative separation-of-function phenotype, the mutant is about 5times less active than the wild type enzyme in cleavage of Holliday junctions, fails to inhibit the dATPase activity of RecA because it does not bind ssDNA
R71A
mutant with a putative separation-of-function phenotype, fails to inhibit the dATPase activity of RecA because it does not bind ssDNA
Y80A
-
naturally occuring mutant showing an an intermediate phenotype. The mutant poorly recognizes and distorts Holliday junctions, and cleaves them with low efficiency. RecUY80A does not inhibit RecA dATPase and strand-exchange activities and it loses structural selectivity for X-shaped structures. Phenotype, detailed overview
D292N
-
1.2fold decrease in binding of four-way DNA junction, 80fold decrease in catalytic activity
D293N
-
1.2fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
D294N
-
1.2fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
E145Q
-
0.85fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
E231A
-
21fold decrease in binding of four-way DNA junction, 4fold decrease in catalytic activity
F79A
-
70fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
K291A
K291R
-
130fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
Q147A
-
7.5fold decrease in binding of four-way DNA junction, 100fold decrease in catalytic activity
R146A
-
70fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
R150A
-
18fold decrease in binding of four-way DNA junction, 70fold decrease in catalytic activity
R231K
-
330fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
D8N
Ceduovirus bIL67
-
mutant defective in one of four conserved residues known to comprise the catalytic site, slightly improved binding to DNA but unable to cleave it
E68G/H136R
-
the mutation affects octamer formation, DNA binding, and the stimulation of RuvB helicase activity
H29R/E40G/E68G/K129E/F140S/S177G/D184N
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
H29R/E40G/Q58R/K129E/F140S/S177G/D184N
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
N73A
-
mutant enzyme with 20% of the wild-type activity at 50 nM protein concentration
N79D/N100D
-
the mutation affects octamer formation, DNA binding and the stimulation of RuvB helicase activity
D132A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D132C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D135N
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D55A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D7C
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
D7N
-
site-directed mutagenesis, inactive mutant, the active-site mutant resolvase holds the arms in a more planar arrangement in EDTA, Ca2+, or Mg2+ conditions
E33A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
E60C
-
site-directed mutagenesis, mutant with reduced activity, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
E60N
-
site-directed mutagenesis, mutant with reduced activity, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
K102A
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
K102R
-
site-directed mutagenesis, mutant enzyme activity with Mg2+ or Mn2+ compared to the wild-type enzyme
R13M
-
mutant is used to include a second methionine in addition to Met-56
Y141A/L142A/N143A/V148A
-
the mutations completely abrogate GEN1 centrosome accumulation
D28N
mutation reduced its ability to cleave Holliday junction DNA
D112A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
D57A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
D68A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
E11A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
E70A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
F103A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F108A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F69A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
F79A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G100A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G60A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
G64A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
G7A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
H87A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
H91A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
K31A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
K72A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
K76A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
L10A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding and cleaving activities
L122A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
L92A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
M8A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
N15A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding and cleaving activities
N5A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding and cleaving activities
Q88A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but no cleaving activities
S54A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding and cleaving activities
T74A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
V46A
-
site-directed mutagenesis, the mutant shows unaltered Holliday junction binding but reduced cleaving activities compared to the wild-type enzyme
Y58A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
Y62A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
Y66A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
G64A
-
site-directed mutagenesis, the mutant shows slightly reduced Holliday junction binding but unaltered cleaving activities compared to the wild-type enzyme
-
H91A
-
site-directed mutagenesis, inactive mutant without Holliday junction binding or cleaving activities
-
K72A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
-
K76A
-
site-directed mutagenesis, the mutant shows reduced Holliday junction binding activity and no cleaving activity
-
A204E
-
the mutant retains some Holliday junction-binding ability, while its cleavage activity is drastically impaired
D116A
-
the mutation completely abolishes Holliday junction cleavage activity, although the binding activity is retained
D118A
-
the mutation completely abolishes Holliday junction cleavage activity, although the binding activity is retained
E175A
-
the mutation completely abolishes Holliday junction cleavage activity, although the binding activity is retained
E258A
-
the mutation completely abolishes Holliday junction cleavage activity, although the binding activity is retained
G200E
-
the mutant retains some Holliday junction-binding ability, while its cleavage activity is drastically impaired
K185D
-
the Holliday junction cleavage activity of the mutant is completely lost
K218D
-
the Holliday junction cleavage activity of the mutant is completely lost
R149D
-
the Holliday junction cleavage activity of the mutant is completely lost
R149D/K185D/K218D/K225D
-
the mutations result in total loss of Holliday junction binding and cleavage activity
R250/K251/K252D
-
the mutations result in total loss of Holliday junction binding and cleavage activity
D33A
DELTA1-5
-
mutation causes a considerable decrease in Hjc-Holliday junction complex formation and cleavage activity
F72A
-
no cleavage of Holliday junction. Mutant enzyme exists as monomer more frequently in solution than as dimer
K30A/K31A
-
mutant enzyme retains proper binding ability to the Holliday junction, little or almost no cleavage activity
K48A
-
no cleavage of Holliday junction. Mutant enzyme exists as monomer more frequently in solution than as dimer
K51A/K52A
-
mutant enzyme retains proper binding ability to the Holliday junction, weak cleavage activity
R3A/K4A
-
mutation reduces the activity by 20fold as compared with the wild-type enzyme. The binding to the Holliday junction is substantially lowered
S30A
-
serine 30 on a flexible loop is catalytically essential, mutants show a decrease in catalytic rate of 3-4 orders of magnitude
S30C
-
serine 30 on a flexible loop is catalytically essential, mutants show a decrease in catalytic rate of 3-4 orders of magnitude
S30T
-
has a slightly higher activity than mutants S30A and S30C but is still severely compromised
D414A/D415A
-
mutations introduce a diagnostic NheI restriction site
DELTA1-35
-
mutant enzyme is not able to resolve the synthetic four-way junction X12
DELTA1-35/CDELTA15
-
mutant enzyme is not able to resolve the synthetic four-way junction X12
E12A
mutant, which has a deficient DNA-cutting but normal DNA-binding activity
S34A
-
the cleavage activity of the phosphorylation-mimic mutant is reduced compared to the wild type enzyme
S34E
-
the cleavage activity of the phosphorylation-mimic mutant is completely lost. The mutant is more resistant to higher doses of cisplatin and UV treatment compared to the wild type enzyme
S9E
-
the cleavage activity of the phosphorylation-mimic mutant is greatly reduced compared to the wild type enzyme
E12A
-
mutant, which has a deficient DNA-cutting but normal DNA-binding activity
-
S34A
-
the cleavage activity of the phosphorylation-mimic mutant is reduced compared to the wild type enzyme
-
S9E
-
the cleavage activity of the phosphorylation-mimic mutant is greatly reduced compared to the wild type enzyme
-
F73A
the mutant shows 50% reduced activity compared to the wild type enzyme
F74A
the mutant shows 30% reduced activity compared to the wild type enzyme
H143D
the mutant shows increased activity compared to the wild type enzyme
M108C
the mutant shows reduced activity compared to the wild type enzyme
P40C
the mutant shows reduced activity compared to the wild type enzyme
R140C
the mutant shows reduced activity compared to the wild type enzyme
T11C
the mutant shows reduced activity compared to the wild type enzyme
Y75A
Y75A/H143D
the mutant shows increased activity compared to the wild type enzyme
F73A
-
the mutant shows 50% reduced activity compared to the wild type enzyme
-
F74A
-
the mutant shows 30% reduced activity compared to the wild type enzyme
-
H143D
-
the mutant shows increased activity compared to the wild type enzyme
-
T11C
-
the mutant shows reduced activity compared to the wild type enzyme
-
Y75A
Y75A/H143D
-
the mutant shows increased activity compared to the wild type enzyme
-
D30N
D81Q
-
mutation eliminates catalytic activity without affecting specific DNA binding
additional information
K291A
-
130fold decrease in binding of four-way DNA junction, 47fold decrease in catalytic activity
K291A
-
30fold decrease in binding of four-way DNA junction, more than 200fold decrease in catalytic activity
D33A
-
mutant enzyme retains proper binding ablity to the Holliday junction
Y75A
the mutant shows about 1.9fold increase of activity compared to the wild type enzyme
Y75A
the mutant shows markedly increased activity compared to the wild type
Y75A
-
the mutant shows about 1.9fold increase of activity compared to the wild type enzyme
-
Y75A
-
the mutant shows markedly increased activity compared to the wild type
-
D30N
-
mutation eliminates catalytic activity without affecting specific DNA binding
additional information
-
gen-1 mutants are defective in DNA damage-dependent cell cycle arrest and apoptosis and in positional cloning of GEN-1, phenotypes, detailed overview
additional information
-
RusA restores viability to polA, dam, and uvrD mutant cells lacking RuvABC
additional information
-
generation of a series of 16 deletion mutants, 9 N- and 7 C-terminal deletion mutants, and 31 point mutants of RecUMge. Deletion of more than 8 amino acid residues renders the mutants inactive, overview
additional information
-
generation of a series of 16 deletion mutants, 9 N- and 7 C-terminal deletion mutants, and 31 point mutants of RecUMge. Deletion of more than 8 amino acid residues renders the mutants inactive, overview
-
additional information
-
construction of mms4DELTA and yen1DELTA deletion mutants, and of the double mutant mms4DELTA/yen1DELTA showing high sensitivity to DNA damage and a specific reduction in crossover events. Genetic interactions of mms4DELTA and yen1DELTA, overview
additional information
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cells that lack Holliday junction resolvase Ydc2 show a significant depletion of mtDNA content. Truncated mutants lacking the SAP motif alone or the whole triple-helix domain are not functional in yeast mitochondria
additional information
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Spcce1:ura4+ insertion mutant strain devoid of the Holliday junction resolvase activity. The majority of mitochondrial DNA from the mutant strain is in an aggregated form apparently due to extensive interlinking of DNA molecules by recombinant junctions. This marked effect on the conformation of mitochondrial DNA results in little or no effect on proliferation or viability of the mutant strain
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 37
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MUS81-EME2 is more stable at 30°C than 37°C, but remains active at 37°C up to 30 min of incubation
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the tetrameric RuvA-Holliday junction complex II is stable in the presence of 750 mM NaCl, whereas the same amount of salt induces more than 80% dissociation of the octameric complex I
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, fraction VI of the Endo X3 purification can be stored after dialysis against buffer E
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-20°C, stable for at least 8 months without measurable loss of activity
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on ice, fraction VI of the Endo X3 purification can be stored for over 1 month, should not be frozen
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
amylose and heparin agarose chromatography of MBP-Rap, Rap29K, Rap-S and Rap17K
Lambdavirus lambda
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carboxy-terminal His-tagged GEN1 is purified using HisTrap column chromatography, heparin column chromatography, ssDNA column chromatography, and MonoS column chromatography
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DEAE-cellulose column chromatography, ammonium sulfate fractionation, Superdex-200 gel filtration, phosphocellulose column chromatography, and hydroxyapatite Bio-Gel HTP column chromatography
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EcME is purified to apparent homogeneity through affinity chromatography, ion exchange, and gel filtration steps
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Endo X3, more than 1325fold, purification beyond fraction V requires addition of pure protein like bovine serum albumin to a final concentration of 0.5 mg/ml
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gel filtration on a Hi-Load (1.6 x 60 cm) Superdex 200 column, 90% purified by SDS-PAGE
highly purified in bacteria, partially purified from the cognate host
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hydroxyapatite column chromatography, MonoQ column chromatography, and Superdex 75 gel filtration
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Ni-NTA column chromatography, HiTrap heparin column chromatography and Superdex 200 gel filtration
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nickel affinity column chromatography and Mono-S column chromatography
overexpressed GST-Mms4/His10-Flag-Mus81 is purified by sequential affinity chromatography from Saccharomyces cerevisiae cells
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partial
phosphocellulose and DNA agarose chromatography, FPLC-gel filtration, 99% purified
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purified to homogeneity
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
amino acids 260-551 for Mus81 and amino acids 244-571 for Eme1 are cloned for expression in Escherichia coli, the complex is named EcME
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C-terminal His-tagged Hjc is overexpressed in Sulfolobus islandicus and coexpression with ATPase SisPINA in Escherichia coli
cloned into the pDuetMxe vector for overexpression in Escherichia coli BL21DE3pLysS cells
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cloning in Escherichia coli strain NEB 5 alpha, and expression in strain NEB Turbo and NEB T7 Express
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cloning of MUS81 and MMS4 into GAL1/10 divergent promotor, 2 micro-based overexpression vector, based on pJN58
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expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expression in Escherichia coli using vector pET19b-Hje with a native N-terminal
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expression of covalently linked endonuclease VII dimers in Escherichia coli BL21
Tequatrovirus T4
expression of MBP-RuvC in Escherichia coli strain ER2508
for expression in Escherichia coli
gene gen-1, DNA and amino acid sequence determination and analysis. GEN-1 assignment and phylogenetic relationships
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gene MG352, expression of wild-type and mutant enzymes as MBP fusion protein sin Escherichia coli strain BL21(DE3)
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gene RecUMge, DNA and amino acid sequence determination and analysis
HJE gene cloned from genomic DNA into the pET19b Escherichia coli expression vector and the recombinant protein is expressed at high levels
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in Escherichia coli strain BL21(DE3)(pLysS) containing the recU overexpression plasmid pCB210
overexpression in Escherichia coli BL21
wild-type ruvAB from Mycobacterium tuberculosis is cloned into pALACE, and the recombinant plasmid (pALACE-ruvAB) is introduced into strain M136 by electroporation to generate the complemented strain Mycobacterium smegmatis M136C
YEN1 coding sequence amplified from BY4741 genomic DNA and cloned into pDONR221. Further subcloning of the wild-type or mutant into pYES-DEST52 or pAG416GPD-ccdB-HA to generate pYES-DEST52-YEN1-V5-6xHis, pAG416GPD-YEN1-3xHA or pAG416GPDYEN1E193A/E195A-3xHA. Coding sequence of Yen1-V5-6xHis amplified from pYES-DEST52-YEN1-V5-6xHis and subcloned into the BamHI and HindIII sites of p416ADH to generate p416ADH-YEN1-V5-6xHis
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
the absence of RuvA and RuvB orthologues in humans suggests that RuvA and RuvB may serve as potential targets for clinically useful small molecule enhancers of moxifloxacin
medicine
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the absence of RuvA and RuvB orthologues in humans suggests that RuvA and RuvB may serve as potential targets for clinically useful small molecule enhancers of moxifloxacin
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Release 2023.1 (January 2023)
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