Promoter analysis of the PHO81 gene encoding a 134 kDa protein bearing ankyrin repeats in the phosphatase regulon of Saccharomyces cerevisiae

• Published in: Molecular & general genetics Vol. 238; no. 3; pp. 444 - 454
• Main Authors: Ogawa, N, Noguchi, K.I, Yamashita, Y, Yasuhara, T, Hayashi, N, Yoshida, K, Oshima, Y
• Format: Journal Article
• Language: English
• Published: Germany 01.04.1993
• Subjects: Acid Phosphatase - genetics; Amino Acid Sequence; amino acid sequences; Ankyrins; Base Sequence; binding proteins; binding sites; Chromosome Mapping; Fungal Proteins - chemistry; Fungal Proteins - genetics; genbank/d13228; gene expression; genetic regulation; Molecular Sequence Data; nucleotide sequences; phosphoric monoester hydrolases; promoter regions; Promoter Regions, Genetic; proteins; regulatory proteins; Regulatory Sequences, Nucleic Acid; Repetitive Sequences, Nucleic Acid; Repressor Proteins; restriction mapping; RNA, Fungal - analysis; RNA, Messenger - analysis; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; structural genes; transcription (genetics)
• ISSN: 0026-8925; 1432-1874
• DOI: 10.1007/bf00292004

The PHO81 gene encoding one of the regulators of the phosphatase regulon in Saccharomyces cerevisiae was mapped 9.8 centimorgans distal from the ser2 locus on the right arm of chromosome VII. Determination of the nucleotide sequence of cloned PHO81 DNA revealed a 3537 bp open reading frame encoding a 134 kDa protein. This protein has six repeats of a 33-amino acid sequence homologous to the ankyrin repeat and an asparagine-rich region. Transcription of PHO81 is activated by Pho4 protein in cooperation with Pho2 (i.e., Bas2/Grf10) protein under the influence of the inorganic phosphate (P(i)) concentration in the medium, through the PHO regulatory system. Major transcription initiation sites of PHO81, determined by primer extension analysis, are at nucleotide positions -66 and -65 relative to the ATG codon. Deletion analysis showed that a 95 bp region from nucleotide position -385 to -291 is essential for response to the P(i) signals. Purified Pho4 protein protected a 19 bp region (positions -350 to -332) in the 95 bp fragment from DNase I digestion in vitro and the protected region includes the core sequence 5'-CACGTG-3', which is also observed in other genes of phosphate metabolism.