2.4.2.9: uracil phosphoribosyltransferase
This is an abbreviated version!
For detailed information about uracil phosphoribosyltransferase, go to the full flat file.
Word Map on EC 2.4.2.9
-
2.4.2.9
-
5-fluorouracil
-
pyrimidine
-
salvage
-
5-fluorocytosine
-
suicide
-
prodrug
-
orotate
-
bystander
-
5-fluorouridine
-
oncolytic
-
markerless
-
phosphoribosyltransferases
-
counterselectable
-
flucytosine
-
4-thiouracil
-
medicine
-
virotherapy
-
cd/5-fc
-
gene-directed
-
synthesis
-
analysis
-
molecular biology
-
pharmacology
- 2.4.2.9
- 5-fluorouracil
- pyrimidine
-
salvage
- 5-fluorocytosine
-
suicide
-
prodrug
- orotate
-
bystander
- 5-fluorouridine
-
oncolytic
-
markerless
-
phosphoribosyltransferases
-
counterselectable
-
flucytosine
- 4-thiouracil
- medicine
-
virotherapy
-
cd/5-fc
-
gene-directed
- synthesis
- analysis
- molecular biology
- pharmacology
Reaction
Synonyms
AFR052C, AFR052Cp, AGOS_AFR052C, AgUPRT, CD-UPRT, cytosine deaminase-uracil phosphoribosyltransferase, cytosine deaminase/uracil phosphoribosyltransferase, More, MtUPRT, phosphoribosyltransferase, uracil, Rv3309c, TtUPRT, TtUPRT3, UK/UPRT1, UMP pyrophosphorylase, UMP:pyrophosphate phosphoribosyltransferase, UPP, UPRT, UPRTase, uracil phosphoribosyl transferase, uridine 5'-phosphate pyrophosphorylase, uridine kinase-like protein (uracil phosphoribosyltransferase), uridine kinase/uracil phosphoribosyltransferase 1, uridine monophosphate pyrophosphorylase, uridylate pyrophosphorylase, uridylic pyrophosphorylase
ECTree
2.4.2.9 uracil phosphoribosyltransferaseAdvanced search results
results (
results (Engineering
Engineering on EC 2.4.2.9 - uracil phosphoribosyltransferase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
P131D
-
site-directed mutagenesis, 2-step mutagenic PCR, exchange of proline in 5-phosphoribose 1-diphosphate binding site, 50-60fold reduction of catalytic rate in both reaction directions, about 100fold increase in KM for uracil, strongly reduced Km for 5-phosphoribose 1-diphosphate
K150A
site-directed mutagenesis, slightly reduced kcat, increased Km for 5-phosphoribose 1-diphosphate in presence of GTP, structural sudy
K59A
site-directed mutagenesis, slightly enhanced kcat, increased Km for 5-phosphoribose 1-diphosphate in presence of GTP, structural sudy
R68A
site-directed mutagenesis, slightly reduced kcat, increased Km for 5-phosphoribose 1-diphosphate in presence of GTP, structural sudy
additional information
additional information
construction of T-DNA insertion enzyme mutants, inability to salvage uracil caused a lightdependent dramatic pale-green to albino phenotype, dwarfism and the inability to produce viable progeny in loss-of-function mutants, plastid biogenesis and starch accumulation are affected in all tissues, with the exception of stomata, phenotypes, overview. A graduation in the tolerance to 5-fluorouracil is observed among the upp(+/) heterozygous plants. Functional complementation of the enzyme-deficient Escherichia coli mutant strain BM604, overview. No significant decrease in UPRT activity is measured in the mutants disrupted at uracil kinase-like proteins UKL loci
additional information
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
additional information
-
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
additional information
-
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
-
additional information
Eremothecium gossypii CBS 109.51
-
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
-
additional information
Eremothecium gossypii FGSC 9923
-
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
-
additional information
Eremothecium gossypii NRRL Y-1056
-
the pyrimidine auxotroph Ashbya gossypii mutant Agura3 shows hypersensitivity to uracil. Uridine is the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Overexpression of the AgUPRT encoding-gene in Ashbya gossypii mutant Agura3 leads to similar growth on rich medium containing 5 mM uracil or uridine
-
additional information
engineering the yeast fluorocytosine deaminase (FCY1) gene by creating a fusion with the bacterial uracil phosphoribosyl transferase (UPP) gene results in a recombinant protein that converts the precursor 5-fluorocytosine (5-FC) into 5-fluorouracyl, a drug used in the treatment of a range of cancers, which triggers DNA and RNA damage. The FCY-UPP gene construct is expressed in specific cell types using enhancer trap lines and promoters, demonstrating that this marker acts in a cell-autonomous manner. It can inactivate slow developmental processes like lateral root formation by targeting pericycle cells. A role for the lateral root cap and the epidermis in controlling root growth, a faster response, is also revealed. The 5-FC precursor acts systemically, as demonstrated by its ability to inhibit stomatal movements when supplied to the roots in combination with a guard cell-specific promoter. The tissular inactivation is reversible, and can therefore be used to synchronize plant responses or to determine cell type-specific functions during different developmental stages
additional information
seven independent upp mutants are constructed and found to excrete low amounts of pyrimidines to the growth medium, pyrimidine-dependent transcription regulation of the biosynthetic pyrimidine pyrR1-B-C-Aa1-Ab1-D-F-E operon is impaired in the upp mutants, overview, model of pyrP pyrimidine-dependent regulation by transcription attenuation
additional information
-
seven independent upp mutants are constructed and found to excrete low amounts of pyrimidines to the growth medium, pyrimidine-dependent transcription regulation of the biosynthetic pyrimidine pyrR1-B-C-Aa1-Ab1-D-F-E operon is impaired in the upp mutants, overview, model of pyrP pyrimidine-dependent regulation by transcription attenuation
additional information
-
seven independent upp mutants are constructed and found to excrete low amounts of pyrimidines to the growth medium, pyrimidine-dependent transcription regulation of the biosynthetic pyrimidine pyrR1-B-C-Aa1-Ab1-D-F-E operon is impaired in the upp mutants, overview, model of pyrP pyrimidine-dependent regulation by transcription attenuation
-
additional information
-
expression of fusion protein cytosine deaminase combined with uracil phosphoribosyl transferase leads to local and distant bystander effects against intraprostatic mouse androgen-refractory prostate, RM1, tumors in immunocompetent mice, CDUPRT-GDEPT expression significantly suppresses the aggressive growth of RM1 prostate tumors and lung pseudo-metastases via immune mechanisms involving necrosis and apoptosis, overview
additional information
generation of an upp knockout strain by allelic replacement (using the pPR27xylE plasmid, which contains a thermosensitive origin of replication, the xylE reporter gene and the sacB counterselectable marker), evaluation of it in infected mice. Knockout and complemented strains are validated by a functional assay of uracil incorporation. Recombinant expression of Mycobacterium smegmatis gene upp in Mycobacterium tuberculosis wild-type strain H37Rv, resulting in strain CP, and in the upp knockout (KO) variant of H37Rv. Swiss male mice are infected intravenously with 5 × 105 colony forming units (CFU) of Mycobacterium tuberculosis H37Rv wild-type, knockout, or CP strains
additional information
-
generation of an upp knockout strain by allelic replacement (using the pPR27xylE plasmid, which contains a thermosensitive origin of replication, the xylE reporter gene and the sacB counterselectable marker), evaluation of it in infected mice. Knockout and complemented strains are validated by a functional assay of uracil incorporation. Recombinant expression of Mycobacterium smegmatis gene upp in Mycobacterium tuberculosis wild-type strain H37Rv, resulting in strain CP, and in the upp knockout (KO) variant of H37Rv. Swiss male mice are infected intravenously with 5 × 105 colony forming units (CFU) of Mycobacterium tuberculosis H37Rv wild-type, knockout, or CP strains
additional information
-
generation of an upp knockout strain by allelic replacement (using the pPR27xylE plasmid, which contains a thermosensitive origin of replication, the xylE reporter gene and the sacB counterselectable marker), evaluation of it in infected mice. Knockout and complemented strains are validated by a functional assay of uracil incorporation. Recombinant expression of Mycobacterium smegmatis gene upp in Mycobacterium tuberculosis wild-type strain H37Rv, resulting in strain CP, and in the upp knockout (KO) variant of H37Rv. Swiss male mice are infected intravenously with 5 × 105 colony forming units (CFU) of Mycobacterium tuberculosis H37Rv wild-type, knockout, or CP strains
-
additional information
-
generation of an upp knockout strain by allelic replacement (using the pPR27xylE plasmid, which contains a thermosensitive origin of replication, the xylE reporter gene and the sacB counterselectable marker), evaluation of it in infected mice. Knockout and complemented strains are validated by a functional assay of uracil incorporation. Recombinant expression of Mycobacterium smegmatis gene upp in Mycobacterium tuberculosis wild-type strain H37Rv, resulting in strain CP, and in the upp knockout (KO) variant of H37Rv. Swiss male mice are infected intravenously with 5 × 105 colony forming units (CFU) of Mycobacterium tuberculosis H37Rv wild-type, knockout, or CP strains
-
additional information
-
sensitivity of cells to 5-FU can be further increased by expression of UPRT, useful in an approach for the management of graft-versus-host disease occurring after allogeneic hematopoietic cell transplantation by genetical engineering of donor T lymphocytes to express a suicide gene, i.e. the Herpes simplex virus thymidine kinase HSV-tk gene, to facilitate negative selection, combination of UPRT expression with expression of cytosine deaminase, an alternative gene for negative selection, converting 5-fluorocytosine to the toxic metabolite 5-fluorouracil, a construct NG/CDiU expressing UPRT and NG/CD, using a bicistronic message, provides the greatest UPRT activity and killing, overview
additional information
covalent immobilization of uracil phosphoribosyltransferase (UPRT) from Thermus thermophilus HB8 onto glutaraldehyde-activated magnetic iron oxide porous microparticles (MTtUPRT). The support is activated by contacting the microspheres with 0.2 ml of 50 mM sodium phosphate buffer, pH 8.5, containing 5% w/v glutaraldehyde during 3 h at 25°C, and then washed thoroughly with distilled water to remove the excess activating agent. Finally, activated microspheres are washed and equilibrated in binding buffer (50 mM potassium phosphate buffer, pH 8.5), shortly prior to use. According to the catalyst load experiments, MTtUPRT3 is selected as optimal biocatalyst. MTtUPRT3 is active and stable in a broad range of temperature (70-100°C) and in the pH interval 6.0-8.0, displaying maximum activity at 100°C and pH 7.0 (activity 968 IU/g support, retaining activity 100%). In addition, MTtUPRT3 can be reused up to 10 times in the synthesis of uridine-5'-monophosphate (UMP), completely stable for 6 cycles. Biocatalyst recycling is studied by employing MTtUPRT3 in consecutive reactions. MTtUPRT3 is successfully applied in the sustainable synthesis of different 5-modified uridine-5'-monophosphates at short times
additional information
-
covalent immobilization of uracil phosphoribosyltransferase (UPRT) from Thermus thermophilus HB8 onto glutaraldehyde-activated magnetic iron oxide porous microparticles (MTtUPRT). The support is activated by contacting the microspheres with 0.2 ml of 50 mM sodium phosphate buffer, pH 8.5, containing 5% w/v glutaraldehyde during 3 h at 25°C, and then washed thoroughly with distilled water to remove the excess activating agent. Finally, activated microspheres are washed and equilibrated in binding buffer (50 mM potassium phosphate buffer, pH 8.5), shortly prior to use. According to the catalyst load experiments, MTtUPRT3 is selected as optimal biocatalyst. MTtUPRT3 is active and stable in a broad range of temperature (70-100°C) and in the pH interval 6.0-8.0, displaying maximum activity at 100°C and pH 7.0 (activity 968 IU/g support, retaining activity 100%). In addition, MTtUPRT3 can be reused up to 10 times in the synthesis of uridine-5'-monophosphate (UMP), completely stable for 6 cycles. Biocatalyst recycling is studied by employing MTtUPRT3 in consecutive reactions. MTtUPRT3 is successfully applied in the sustainable synthesis of different 5-modified uridine-5'-monophosphates at short times
