Reciprocal regulation between Slt2 MAPK and isoforms of Msg5 dual-specificity protein phosphatase modulates the yeast cell integrity pathway.
Dual-specificity protein phosphatases (DSPs) are involved in the negative regulation of mitogen-activated protein kinases (MAPKs) by dephosphorylating both threonine- and tyrosine-conserved residues located at the activation loop. Here we show that Msg5 DSP activity is essential for maintaining a low level of signaling through the cell integrity pathway in Saccharomyces cerevisiae. Consistent with a role of this phosphatase on cell wall physiology, cells lacking Msg5 displayed an increased sensitivity to the cell wall-interfering compound Congo Red. We have observed that the N-terminal non-catalytic region of this phosphatase was responsible for binding to the kinase domain of Slt2, the MAPK that operates in this pathway. In vivo and in vitro experiments revealed that both proteins act on each other. Msg5 bound and dephosphorylated activated Slt2. Reciprocally, Slt2 phosphorylated Msg5 as a consequence of the activation of the cell integrity pathway. In addition, alternative use of translation initiation sites at MSG5 resulted in two protein forms that are functional on Slt2 and became equally phosphorylated following activation of this MAPK. Under activating conditions, a decrease in the affinity between Msg5 and Slt2 was observed, leading us to suggest that the mechanism by which Slt2 controls the action of Msg5 was via the modulation of protein-protein interactions. Our results indicate the existence of posttranscriptional mechanisms of regulation of DSPs in yeast and provide new insights into the negative control of the cell integrity pathway.
Flández
Marta
M
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Pza Ramón y Cajal s/n., 28040 Madrid, Spain.
Cosano
Inmaculada C
IC
Nombela
César
C
Martín
Humberto
H
Molina
María
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; D029701:Saccharomyces cerevisiae Proteins; D020928:Mitogen-Activated Protein Kinases; C085791:MSG5 protein, S cerevisiae; D017027:Protein Tyrosine Phosphatases