Isolation of phosphoribosylpyrophosphate synthetase (PRS1) gene from Candida albicans.

We have isolated a 3.7 kb EcoR1 fragment from a genomic library of Candida albicans which displayed a 65% level of identity with the PRS gene family (PRS) of Saccharomyces cerevisiae. The PRS gene encodes a phosphoribosylpyrophosphate (PRPP) synthetase of S. cerevisiae, which catalyses the synthesis of purines, pyrimidines, and amino acids such as histidine and tryptophan. By Northern analyses, we observed that the entire 3.7 kb EcoR1 fragment as well as 1.1 kb KpnI-SacI internal fragment of the 3.7 kb EcoR1 fragment hybridized to the same 1.4 kb transcript. An internal 2.6 kb KpnI fragment was subcloned and sequenced. A deduced sequence of 321 amino acids representing a polypeptide of 35.2 kDa was determined. A FASTA search indicated that the C. albicans PRS (Ca PRS1) had an overall homology at the amino acid level of 91% with the S. cerevisiae PRS3. Putative transcriptional start and termination sequences as well as a cation-binding, PRPP synthetase signature sequence were identified. Ca PRS1 was localized to chromosome 2 of the C. albicans genome. Low stringency hybridizations indicates that the organism may possess multiple PRS genes. The function of these genes in nitrogen signaling is discussed.