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A Reb1p‐binding site is required for efficient activation of the yeast RAP1 gene, but multiple binding sites for Rap1p are not essential
Author(s) -
Graham Ian R.,
Chambers Alistair
Publication year - 1994
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1994.tb01081.x
Subject(s) - rap1 , biology , binding site , transcription factor , gene , saccharomyces cerevisiae , microbiology and biotechnology , promoter , transcription (linguistics) , dna binding site , telomere binding protein , dna , genetics , gene expression , dna binding protein , linguistics , philosophy
Summary The Saccharomyces cersvislae RAP1 protein (Rap1p) is a key multifunctional transcription factor. Using gel retardation analysis, four binding sites for RAP1 p have been identified within the promoter of the RAP1 gene. These sites are located downstream of a binding site for the transcription factor Reb1p. The Reb1p site and an associated AT‐rich region are important for transcriptional activation, but deletion of three of the RAP1 p‐binding sites had little effect on promoter activity. The activity of the RAP1 promoter has been analysed in a yeast strain (YDS410) that contains a temperature‐sensitive mutation In the RAP1 gene. This mutation renders the DNA‐binding activity of Rapip temperature dependent. When YDS410 was grown at a semi‐permissive temperature (30°C), the activity of the RAP1 promoter increased by approximately 170%, compared with the same strain grown at the permissive temperature (25°C). A RAP1 promoter in which three of the four RAP1 p‐binding sites had been deleted, showed only a small increase in activity in the same experiment. These data confirm that Rap1p is not required for activation of the RAP1 gene, and suggest a role for Rap1p In negative auto‐regulation.