Altered phosphotransfer in an activated mutant of the Saccharomyces cerevisiae two-component osmosensor Sln1p.
The SLN1 two-component signaling pathway of Saccharomyces cerevisiae utilizes a multistep phosphorelay mechanism to control osmotic stress responses via the HOG1 mitogen-activated protein kinase pathway and the transcription factor Skn7p. Sln1p consists of a sensor kinase module that undergoes histidine autophosphorylation and a receiver module that autocatalytically transfers the phosphoryl group from histidine to aspartate. The Sln1p aspartyl phosphate is then transferred to Ypd1p, which in turn transfers the phosphoryl group to a conserved aspartate on one of two response regulators, Ssk1p and Skn7p. Activated alleles of SLN1 (sln1*) were previously identified that appear to increase the level of phosphorylation of downstream targets Ssk1p and Skn7p. In principle, the phenotype of sln1* alleles could arise from an increase in autophosphorylation or phosphotransfer activities or a decrease in an intrinsic or extrinsic dephosphorylation activity. Genetic analysis of the activated mutants has been unable to distinguish between these possibilities. In this report, we address this issue by analyzing phosphorelay and phosphohydrolysis reactions involving the Sln1p-associated receiver. The results are consistent with a model in which the activated phenotype of the sln1* allele, sln-22, arises from a shift in the phosphotransfer equilibrium from Sln1p to Ypd1p, rather than from impaired dephosphorylation of the system in response to osmotic stress.
Ault
A D
AD
Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242, USA.
Fassler
J S
JS
Deschenes
R J
RJ
D004268:DNA-Binding Proteins / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008957:Models, Genetic; D009154:Mutation; D009997:Osmotic Pressure; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C083962:SKN7 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D011494:Protein Kinases; C102035:YPD1 protein, S cerevisiae; C083749:SLN1 protein, S cerevisiae