THE PPZ PROTEIN PHOSPHATASES ARE INVOLVED IN THE MAINTENANCE OF OSMOTIC STABILITY OF YEAST-CELLS

We have recently reported the existence in the yeast Saccharomyces cerevisiae of a gene named PPZ1, encoding a novel Ser/Thr phosphatase characterized by a large, Ser-rich amino-terminal extension, and suggested the existence of a related gene product that could have overlapping functions. We have now amplified by polymerase chain reaction techniques a genomic fragment of about 600 bp corresponding to this second gene (PPZ2). This fragment hybridizes to an mRNA of about the same size as the PPZ1 message but the amount of PPZ2 mRNA peaks at the stationary phase, when almost no PPZ1 mRNA is found. The PPZ2 fragment was interrupted in vitro and used to transform diploid heterozygous ppz1 PPZ2 cells. Haploid cells carrying the double mutation ppz1 ppz2 were unable to grow in the presence of 5 mM caffeine. However, the mutants did survive when osmotically stabilized in the presence of 1 M sorbitol. The evidence obtained suggests that PPZ1 and PPZ2 may be structurally and functionally related and points to an involvement of these phosphatases in functions related to the maintenance of cell integrity.