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URA5  -  orotate phosphoribosyltransferase URA5

Saccharomyces cerevisiae S288c

Synonyms: OPRT 1, OPRTase 1, Orotate phosphoribosyltransferase 1, PYR5, YM8339.13, ...
 
 
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Disease relevance of URA5

 

High impact information on URA5

 

Biological context of URA5

 

Associations of URA5 with chemical compounds

  • Uracil-requiring mutants of K. lactis CBS 683 were selected on media containing 5-fluoro-orotic acid and used as recipients in transformation experiments using K. lactis URA5 as the selectable marker, thereby proving functionality of the cloned gene [11].
  • URA5 genes encode orotidine-5'-monophosphate pyrophosphorylase (OMPpase), an enzyme involved in pyrimidine biosynthesis [12].
  • Isolation and confirmation of function of the Coccidioides immitis URA5 (orotate phosphoribosyl transferase) gene [1].
  • Several strategies commonly used in Saccharomyces cerevisiae and other eukaryotes were unsuccessful, due to the frequent occurrence of ectopic integration, linear plasmid formation, and spontaneous resistance to 5-fluoroorotic acid, which is a selective agent for URA5 gene inactivation [12].
  • We cloned the Histoplasma capsulatum URA5 gene (URA5Hc) by using a probe generated by PCR with inosine-rich primers based on relatively conserved sequences in OMPpases from other organisms [12].
 

Other interactions of URA5

 

Analytical, diagnostic and therapeutic context of URA5

  • Transformation of the wdura5Delta mutant to prototrophy was accomplished by electroporation of Wd yeast cells with the Ci URA5 gene [1].
  • Northern-blot hybridization revealed the URA5 transcript to be approximately 0.94 kb [13].
  • Isothermal titration calorimetry (ITC) experiments revealed 1:1 stoichiometries for both OMP and Mg(2+)-PRPP with OPRTase yielding K(d) values of 0.68 and 10 microM, respectively [5].
  • Radiolabeled orotidine 5'-phosphate (OMP), generated in situ from [7-(14)C]-orotate and alpha-d-5-phoshorylribose 1-diphosphate (PRPP), binds tightly enough to OPRTase (a dimer composed of identical subunits) that the complex survives gel-filtration chromatography [5].
  • Both orotate phosphoribosyltransferase (OPRTase) and hypoxanthine/guanine phosphoribosyltransferase (HGPRTase) have been purified from Baker's yeast and analyzed kinetically using a modification of published HPLC procedures [14].

References

  1. Isolation and confirmation of function of the Coccidioides immitis URA5 (orotate phosphoribosyl transferase) gene. Yu, J.J., Zheng, L., Thomas, P.W., Szaniszlo, P.J., Cole, G.T. Gene (1999) [Pubmed]
  2. Kinetic mechanism of orotate phosphoribosyltransferase from Salmonella typhimurium. Bhatia, M.B., Vinitsky, A., Grubmeyer, C. Biochemistry (1990) [Pubmed]
  3. Study of the kinetic and physical properties of the orotidine-5'-monophosphate decarboxylase domain from mouse UMP synthase produced in Saccharomyces cerevisiae. Langdon, S.D., Jones, M.E. J. Biol. Chem. (1987) [Pubmed]
  4. Kinetic analysis of nicotinate phosphoribosyltransferase from yeast using high pressure liquid chromatography. Hanna, L.S., Hess, S.L., Sloan, D.L. J. Biol. Chem. (1983) [Pubmed]
  5. Half-of-sites binding of orotidine 5'-phosphate and alpha-D-5-phosphorylribose 1-diphosphate to orotate phosphoribosyltransferase from Saccharomyces cerevisiae supports a novel variant of the Theorell-Chance mechanism with alternating site catalysis. McClard, R.W., Holets, E.A., MacKinnon, A.L., Witte, J.F. Biochemistry (2006) [Pubmed]
  6. A novel enzyme complex of orotate phosphoribosyltransferase and orotidine 5'-monophosphate decarboxylase in human malaria parasite Plasmodium falciparum: physical association, kinetics, and inhibition characterization. Krungkrai, S.R., DelFraino, B.J., Smiley, J.A., Prapunwattana, P., Mitamura, T., Horii, T., Krungkrai, J. Biochemistry (2005) [Pubmed]
  7. Molecular and genetic analysis of URA5 transformants of Cryptococcus neoformans. Varma, A., Edman, J.C., Kwon-Chung, K.J. Infect. Immun. (1992) [Pubmed]
  8. Cloning and sequencing of URA10, a second gene encoding orotate phosphoribosyl transferase in Saccharomyces cerevisiae. de Montigny, J., Kern, L., Hubert, J.C., Lacroute, F. Curr. Genet. (1990) [Pubmed]
  9. A counter-selectable marker for genetic transformation of the yeast Schwanniomyces alluvius. Dave, M.N., Chattoo, B.B. Appl. Microbiol. Biotechnol. (1997) [Pubmed]
  10. Sequence analysis of the DdPYR5-6 gene coding for UMP synthase in Dictyostelium discoideum and comparison with orotate phosphoribosyl transferases and OMP decarboxylases. Jacquet, M., Guilbaud, R., Garreau, H. Mol. Gen. Genet. (1988) [Pubmed]
  11. The URA5 gene encoding orotate-phosphoribosyl transferase of the yeast Kluyveromyces lactis: cloning, sequencing and use as a selectable marker. Bai, X., Larsen, M., Meinhardt, F. Yeast (1999) [Pubmed]
  12. Rare homologous gene targeting in Histoplasma capsulatum: disruption of the URA5Hc gene by allelic replacement. Woods, J.P., Retallack, D.M., Heinecke, E.L., Goldman, W.E. J. Bacteriol. (1998) [Pubmed]
  13. Cloning and sequencing of the URA5 gene from the yeast Yarrowia lipolytica. Sánchez, M., Prado, M., Iglesias, F.J., Domínguez, A. Yeast (1995) [Pubmed]
  14. Enzymatic kinetic analyses that employ high-performance liquid chromatography. Competition between orotate- and hypoxanthine/guanine-phosphoribosyltransferases for a common substrate. Chung, S.H., Sloan, D.L. J. Chromatogr. (1986) [Pubmed]
 
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