Cloning and sequencing of URA10, a second gene encoding orotate phosphoribosyl transferase in Saccharomyces cerevisiae |
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Authors: | J de Montigny L Kern J -C -C Hubert F Lacroute |
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Institution: | (1) Laboratoire de génétique physiologique, I.B.M.C. du C.N.R.S., 15, rue Descartes, F-67084 Strasbourg Cedex, France;(2) Present address: Laboratoire de Microbiologie, Université Louis Pasteur, 4 rue Blaise Pascal, F-67070 Strasbourg Cedex, France;(3) Present address: Centre de Génétique Moléculaire du C.N.R.S., F-91191 Gif-sur-Yvette Cedex, France |
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Abstract: | Summary Orotate phosphoribosyl transferase (OPRTase) catalyses the transformation of orotate to OMP in the pyrimidine pathway. In the yeast Saccharomyces cerevisiae, the URA5 gene is known to encode this enzyme activity. In this paper we present the cloning and sequencing of a yeast gene, named URA10, encoding a second OPRTase enzyme. Comparison of the predicted amino acid sequences between URA5 and URA10 genes shows more than 75% similarity. These sequences have also been compared to those of Escherichia coli, Podospora anserina, Sordaria macrospora and Dictyostelium discoideum. Remarkable similarities in the primary structure of these proteins have been found. Gene disruption experiments revealed that URA10 gene expression is responsible for the leaky phenotype of a ura5 mutant. Assays of OPRTase activity in extracts from ura5 and ura10 mutants indicate that the URA10 product contributes only 20% of the total activity found in wild type cells. |
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Keywords: | Saccharomyces cerevisiae Orotate phosphoribosyl transferase Nucleotide sequence-5-phosphoribosyl 1-pyrophosphate (5PRPP) |
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