The interaction of Slt2 MAP kinase with Knr4 is necessary for signalling through the cell wall integrity pathway in Saccharomyces cerevisiae.
In budding yeast, PKC1 plays an essential role in cell integrity and proliferation through a linear MAP (Mitogen Activated Protein) kinase phosphorylation cascade, which ends up with the activation of the Slt2-MAP kinase by dual phosphorylation on two conserved threonine and tyrosine residues. In this phosphorylated form, Slt2p kinase activates by phosphorylation at least two known downstream targets: Rlm1p, which is implicated in the expression of cell wall-related genes, and SBF, required for transcription activation of cell cycle-regulated genes at the G1 to S transition. In this paper, we demonstrate by two-hybrid, in vitro immunoprecipitation and tandem affinity purification (TAP) methods that Knr4p physically interacts with Slt2p. Moreover, we show that the absence of Knr4p alters proper signalling of Slt2p to its two known downstream targets. In a knr4 null mutant, the SLT2-dependent activation of Rlm1p is strongly reduced and the transcriptional activity of Rlm1p is decreased, although the phosphorylated form of Slt2p is more abundant than in wild-type cells. On the contrary, SBF is abnormally activated in this mutant, as shown by a more abundant phosphorylated form of Swi6p, by higher beta-galactosidase levels from a SCB-lacZ gene fusion, and by deregulation of the cyclic behaviour of several cell cycle-regulated genes. These results, taken together with our recent finding that Bck2p requires Knr4p to activate additively with Cln3-Cdc28p SBF target genes, lead to a model in which Knr4p is involved in co-ordinating the Slt2p-mediated cell wall integrity pathway with progression of the cell cycle.
Martin-Yken
Helene
H
Centre de Bioingenierie Gilbert Durand, UMR-CNRS 5504, UR-INRA 792, 135, avenue de Rangueil, F-31077, Toulouse, France.
Dagkessamanskaia
Adilia
A
Basmaji
Fadi
F
Lagorce
Arnaud
A
Francois
Jean
J
D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D002473:Cell Wall / Q000378:metabolism*; D017930:Genes, Reporter; D026161:MADS Domain Proteins; D046911:Macromolecular Substances; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques
D026161:MADS Domain Proteins; D046911:Macromolecular Substances; C095725:RLM1 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; C085451:SMI1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D020928:Mitogen-Activated Protein Kinases; C072724:SLT2 protein, S cerevisiae