Nucleic Acids Research, 1993, Vol. 21, No. 8 1975-1982
© 1993
MOLECULAR BIOLOGY |
Molecular analysis of the PHO81 gene of Saccharomyces cerevisiae
1Department of Microbiology and Immunology, Hahnemann University School of Medicine Philadelphia, PA 19102 2Department of Biological Sciences, Ohio University Athens, OH 45701, USA
* To whom correspondence should be addressed
Received October 5, 1992. Revised March 5, 1993. Accepted March 5, 1993.
The PH081 gene product is a positive regulatory factor required for the synthesis of the phosphate represslble acid phosphatase (encoded by the PH05 gene) in Saccharomyces cerevlslae. Genetic analysis has suggested that PH081 may be the signal acceptor molecule; however, the biochemical function of the PHO81 gene product is not known. We have cloned the PHO81 gene and sequenced the promoter. A PHO81-LacZ fusion was shown to be a valid reporter since Its expression is regulated by the level of inorganic phosphate and is controlled by the same regulatory factors that regulate PHO5 expression. To elucidate the mechanism by which PHO81 functions, we have isolated and cloned dominant mutations in the PHO81 gene which confer constitutive synthesis of add phosphatase. We have demonstrated that overexpression of the negative regulatory factor, PHO80, but not the negative regulatory factor PHO85, partially blocks the constitutive acid phosphatase synthesis in a strain containing a dominant constitutive allele of PHO81. This suggests that PHO81 may function by interacting with PHO80 or that these molecules compete for the same target.
+ Present address: The Wistar Institute, University of Pennsylvania, Philadelphia, PA 19104, USA
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