Regulation of the Saccharomyces cerevisiae Slt2 Kinase Pathway by the Stress-inducible Sdp1 Dual Specificity Phosphatase*

The Slt2/Mpk1 mitogen-activated protein kinase (MAPK) cell integrity pathway is involved in maintenance of cell shape and integrity during vegetative growth and mating inSaccharomyces cerevisiae. Slt2 is activated by dual phosphorylation of a threonine and tyrosine residue in response to several environmental stresses that perturb cell integrity. Negative regulation of Slt2 is achieved via dephosphorylation by two protein-tyrosine phosphatases, Ptp2 and Ptp3, and a dual specificity phosphatase, Msg5. In this study, we provide genetic and biochemical evidence that the stress-inducible dual specificity phosphatase, Sdp1, negatively regulates Slt2 by direct dephosphorylation. Deletion ofSDP1 exacerbated growth defects due to overexpression of Mkk1p386, a constitutively active mutant of Slt2 MAPK kinase, whereas overexpression of Sdp1 suppressed lethality caused by Mkk1p386 overexpression. The heat shock-induced phosphorylation level of Slt2 was elevated in an sdp1Δstrain compared with that of the wild type, and heat shock-activated phospho-Slt2 was dephosphorylated by recombinant Sdp1 in vitro. Under normal growth conditions, an Sdp1-GFP fusion protein was localized to both the nucleus and cytoplasm. However, the Sdp1-GFP protein translocated to punctate spots throughout the cell after heat shock. SDP1 transcription was induced by several stress conditions in an Msn2/4-dependent manner but independent of the Rlm1 transcription factor, a downstream target activated by Slt2. Induction of SLT2 by high osmolarity was dependent on Rlm1 transcription factor and Hog1 kinase, suggesting cross-talk between Slt2 and Hog1 MAPK pathways. These studies demonstrate regulation of Slt2 activity and gene expression in coordination with other stress signaling pathways.

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