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J Biol Chem, Vol. 275, Issue 2, 1511-1519, January 14, 2000
From the Departamento de Microbiología II, Facultad de
Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y
Cajal s/n, 28040 Madrid, Spain
Signal transduction mediated by the
mitogen-activated protein kinase (MAPK) Slt2 pathway is essential to
maintain the cell wall integrity in Saccharomyces
cerevisiae. Stimulation of MAPK pathways results in activation by
phosphorylation of conserved threonine and tyrosine residues of MAPKs.
We have used an antibody that specifically recognizes dually
phosphorylated Slt2 to gain insight into the activation and modulation
of signaling through the cell integrity pathway. We show that caffeine
and vanadate activate this pathway in the absence of osmotic
stabilization. The lack of the putative cell surface sensor Mid2
prevents vanadate- but not caffeine-induced Slt2 phosphorylation.
Disruption of the Rho1-GTPase-activating protein genes SAC7
and BEM2 leads to constitutive Slt2 activation, indicating
their involvement as negative regulators of the pathway. MAPK kinases
also seem to participate in signaling regulation, Mkk1 playing a
greater role than Mkk2 in signal transmission to Slt2. Additionally,
one of the phosphatases involved in Slt2 dephosphorylation is likely to
be the dual specificity phosphatase Msg5, since overexpression of
MSG5 in a sac7
Regulatory Mechanisms for Modulation of Signaling through the
Cell Integrity Slt2-mediated Pathway in Saccharomyces
cerevisiae*
,
mutant eliminates the high
Slt2 phosphorylation, and disruption of MSG5 in wild type
cells results in increased phospho-Slt2 levels. These data present the
first evidence for a negative regulation of the cell integrity pathway.
*
This work was supported by Grant BIO98-0726 from CICYT
(Ministerio de Educación y Cultura, Spain).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a predoctoral fellowship from the Ministerio de
Educación y Cultura of Spain.
§
Recipient of a predoctoral fellowship from the Comunidad
Autónoma de Madrid.
¶
To whom correspondence should be addressed. Tel.: 34 91 3941748; Fax: 34 91 3941745; E-mail: molmifa@eucmax.sim.ucm.es.
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