Summary
Yeast URA2 encodes a multifunctional carbamoyl phosphate synthetase-aspartate transcarbamylase of 220,000 molecular weight. We determined the nucleotide sequence of the 5′ proximal part of the gene which is responsible for the glutamine amide transfer function of the carbamoyl phosphate synthetase activity. Alignment of the enzyme sequence derived from URA2 with sequences from Escherichia coli carA carB and yeast arginine-specific CPA1 CPA2 indicates that monofunctional and bifunctional carbamoyl phosphate synthetases are probably homologous. The URA2-derived enzyme organization is NH2-carbamoyl phosphate synthetase-aspartate transcarbamylase-CO2H.
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Amuro N, Paluh JL, Zalkin H (1985) Replacement by site-directed mutagenesis indicates a role for histidine 170 in the glutamine amide transfer function of anthranilate synthase. J Biol Chem 260:14844–14849
Anderson PM, Meister M (1965) Evidence for an active form of carbon dioxide in the reaction catalyzed by Escherichia coli. Biochemistry 4:2803–2809
Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031
Biggin MD, Gibson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965
Chou P, Fasman GD (1978) Prediction of the secondary structure of proteins from their amino acid sequences. Advances Enzymol 47:45–147
Denis-Duphil M, Lacroute F (1971) Fine structure of the URA2 locus in Saccharomyces cerevisiae: I. In vivo complementation studies. Mol Gen Genet 112:354–364
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Hall MN, Hereford L, Herskowitz (1984) Targeting of E. coli β-galactosidase to the nucleus in yeast. Cell 36:1057–1065
Hopp TP, Woods KR (1981) Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci USA 78:3824–3828
Jarry B, Falk DR (1974) Functional diversity within the rudimentary locus of Drosophila melanogaster. Mol Gen Genet 135:113–122
Jones ME (1965) Amino acid metabolism. Annu Rev Biochem 34:381–418
Jones ME (1980) Pyrimidine nucleotide biosynthesis in animals: genes, enzymes and regulation of UMP biosynthesis. Annu Rev Biochem 49:253–279
Kaplan JB, Nichols BP (1983) Nucleotide sequence of Escherichia coli pabA and its evolutionary relationship to trp(G)D. J Mol Biol 168:451–468
Kawamura M, Keim PS, Goto Y, Zalkin H, Heinrikson RL (1978) Anthranilate synthetase component II from Pseudomonas putida. J Biol Chem 253:4659–4668
Kosak M (1981) Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res 9:5233–5252
Kozak M (1984) Selection of initiation sites eucaryotic ribosomes: effect of inserting AUG upstream from the coding sequence for preproinsulin. Nucleic Acids Res 12:3873–3893
Lacroute F (1968) Regulation of pyrimidine biosynthesis in Saccharomyces cerevisiae. J Bacteriol 95:824–832
Lacroute F, Pierard A, Grenson M, Wiame JM (1965) The biosynthesis of carbamylphosphate in Saccharomyces cerevisiae. J Gen Microbiol 40:127–142
Lusty CJ, Widgren EE, Broglic KE, Nyunoya H (1983) Yeast carbamoyl phosphate synthetase: structure of the yeast gene and homology to Escherichia coli carbamoyl phosphate synthetase. J Biol Chem 258:14466–14477
Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321
Mori M, Tatibana M (1978) A multienzyme complex of carbamoyl phosphate synthetase (glutamine): aspartate carbamoyltransferase: dihydroorotase (rat ascites hepatoma cells and rat liver). Methods Enzymol 51:111–121
Nagy M, Laporte J, Penverne B, Herve G (1982) Nuclear localization of aspartate transcarbamoylase in Saccharomyces cerevisiae. J Cell Biol 92:790–794
Nichols BP, Miozzari GF, vanCleemput M, Bennett GN, Yanofsky C (1980) Nucleotide sequences of the trpG regions of Escherichia coli, Shigella dysenteria, Salmonella typhimurium and Serratia marcescens. J Mol Biol 142:503–517
Nyunoya H, Lusty CJ (1984) Sequence of the small subunit of yeast carbamyl phosphate synthetase and identification of its catalytic domain. J Biol Chem 259:9790–9798
Padgett RA, Wahl GM, Stark GR (1982) Structure of the gene for CAD, the multifunctional protein that initiates UMP synthesis in syrian hamster cells. Mol Cell Biol 2:293–301
Pierard A, Schröter B (1978) Structure-function relationships in the arginine pathway carbamoyl phosphate synthase of Saccharomyces cerevisiae. J Bacteriol 134:167–176
Pierard A, Grenson M, Glansdorff N, Wiame JM (1973) A comparison of the organization of carbamyl-phosphate synthesis in Saccharomyces cerevisiae and Escherichia coli, based on genetical and biochemical evidences. In: Prusiner S, Stadtman ER (eds) The enzymes of glutamine metabolism. Academic Press, New York, p 483
Pierard A, Glansdorff N, Mergeay M, Wiame JM (1965) Control of the biosynthesis of carbamoyl phosphate in Escherichia coli. J Mol Biol 14:23–36
Piette J, Nyunoya H, Lusty CJ, Cunin R, Weyens G, Crabeel M, Charlier D, Glansdorff N, Pierard A (1984) DNA sequence of the carA gene and the control region of carAB; tandem promoters, respectively controlled by arginine and the pyrimidines, regulate the synthesis of carbamyol-phosphate synthetase in Escherichia coli K12. Proc Natl Acad Sci USA 81:4134–4138
Potier S, Souciet JL, Lacroute F (1987) Correlation between restriction map, genetic map, and catalytic function in the gene complex URA2. Mol Gen Genet (to appear)
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schechtman MG, Yanofsky C (1983) Structure of the trifunctional trp1 gene from Neurospora crassa and its aberrant expression in Escherichia coli. J Mol Appl Genet 2:83–99
Segraves WA, Louis C, Schedl P, Jarry B (1983) Isolation of the rudimentary locus of Drosophila melanogaster. Mol Gen Genet 189:34–40
Silver PA, Keegan LP, Ptashne M (1984) Amino terminus of the yeast GAL4 gene product is sufficient for nuclear localization. Proc Natl Acad Sci USA 81:5951–5955
Smith M, Leung DW, Gillam S, Astell CR, Montgomery DL, Hall BD (1979) Sequence of the gene for iso-1-cytochrome c in Saccharomyces cerevisiae. Cell 16:753–761
Souciet JL, Hubert JC, Lacroute F (1982) Cloning and restriction mapping of the yeast URA2 gene coding for the carbamoyl phosphate synthetase-aspartate transcarbamylase complex. Mol Gen Genet 186:385–390
Tiedeman AA, Smith JM, Zalkin H (1985) Nucleotide sequence of the guaA gene encoding GMP synthetase of Escherichia coli K12. J Biol Chem 260:8676–8679
Weng M, Makaroff CA, Zalkin H (1985) Nucleotide sequence of Escherichia coli pyrG encoding CTP synthetase. J Biol Chem 261:5568–5574
Werner M, Feller A, Pierard A (1984) Nucleotide sequence of yeast CPA1 encoding the small subunit arginine-pathway carbamoyl phosphate synthetase. Eur J Biochem 146:371–381
Zalkin H, Yanofsky C (1982) Yeast gene TRP5: structure, function, regulation. J Biol Chem 257:1491–1500
Zalkin H, Paluh JL, van Cleemput M, Moye WS, Yanofsky C (1984) Nucleotide sequence of Saccharomyces cerevisiae genes TRP2 and TRP3 encoding bifunctional anthranilate synthase: indole-3-glycerol phosphate synthase. J Biol Chem 259:3985–3992
Zalkin H, Argos P, Narayana SVL, Tiedeman AA, Smith JM (1985) Identification of a trpG-related glutamine amide transfer domain in Escherichia coli GMP synthetase. J Biol Chem 20:3350–3354
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Communicated by C.P. Hollenberg
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Souciet, JL., Potier, S., Hubert, JC. et al. Nucleotide sequence of the pyrimidine specific carbamoyl phosphate synthetase, a part of the yeast multifunctional protein encoded by the URA2 gene. Mol Gen Genet 207, 314–319 (1987). https://doi.org/10.1007/BF00331595
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DOI: https://doi.org/10.1007/BF00331595