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Literature summary for 2.7.7.B22 extracted from

  • Chiruvella, K.K.; Rajaei, N.; Jonna, V.R.; Hofer, A.; Astroem, S.U.
    Biochemical characterization of Kat1 a domesticated hAT-transposase that induces DNA hairpin formation and MAT-switching (2016), Sci. Rep., 6, 21671 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.B22

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Kluyveromyces lactis

Protein Variants

Protein Variants Comment Organism
C130A/C133A mutant losES detectable DNA binding Kluyveromyces lactis
C130A/C133A mutations in the BED zinc-finger domain, result in an unspecific nuclease activity Kluyveromyces lactis
C402A/H405A mutant binds DNA normally and is critical for hairpin formation Kluyveromyces lactis
C402A/H405A mutant completely blocks hairpinning and switching, but still generates nicks in the DNA Kluyveromyces lactis
D310A mutation in the catalytic DDE-motif, catalytically inactive Kluyveromyces lactis
D377A mutation in the catalytic DDE-motif, catalytically inactive Kluyveromyces lactis
E895A mutation in the catalytic DDE-motif, catalytically inactive Kluyveromyces lactis
additional information mutations of predicted catalytic residues abolish both DNA cleavage and strand-transfer. Kat1 mutants defective for cleavage in vitro are also defective for mating-type switching Kluyveromyces lactis
W576A mutation predicted to be impaired for hairpin formation, mutant is active for DNA cleavage and supports wild type levels of mating-type switching Kluyveromyces lactis

Organism

Organism UniProt Comment Textmining
Kluyveromyces lactis
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information incubation of Kat1 and a 197 bp duplex corresponding to TIR-R with 48 bp from the transposon and 149 bp flanking DNA. First Kat1 cleaves the transposon end, generating a free 3' hydroxyl. Joining of one 3'-OH to the target generates a nicked plasmid or single end-joining (SEJ) product. If two 3'-OHs join the target at complementary positions on opposite strands, the plasmid is linearized generating a double end-joining (DEJ) product. Kat1 displays target joining, generating both SEJ- and DEJ-products Kluyveromyces lactis ?
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additional information incubation of Kat1 with a 40 bp duplex representing imperfect inverted repeat TIR-R and including 30 bp from the transposon end and 10 bp from the flanking DNA. Kat1 cleaves the upper strand primarily at GTATA*C and to less extent at GTAT*AC. In the reaction using 3' end-labeling of the upper strand a product of 23 nts is observed. The products formed from the lower strand are hairpin-capped. No substrate: a 40 bp duplex representing imperfect inverted repeat TIR-L Kluyveromyces lactis ?
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Synonyms

Synonyms Comment Organism
hAT-transposase 1
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 Kluyveromyces lactis
Kat1
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 Kluyveromyces lactis

General Information

General Information Comment Organism
physiological function Kat1 promotes joining of the transposon end into a target DNA molecule in vitro. Kat1 can form hexamers when complexed with DNA. Kat1 binds DNA non-specifically and the DNA interaction is dependent on a Zn2+-finger motif in its N-terminus Kluyveromyces lactis