Abstract
The protein kinase C (PKC1) pathway is essential for maintaining cell integrity in yeast. Here it is shown that various forms of cell wall damage result in activation of the downstream MAP kinase Slt2/Mpk1. Several cell wall mutants displayed enhanced FKS2-lacZ expression, a known output of Slt2 activation. A similar response was obtained with wild-type cells grown in the presence of the cell wall perturbants Calcofluor white and Zymolyase. Upregulation of FKS2-lacZ in response to sublethal concentrations of these agents fully depended on the presence of Slt2. The same cell wall stress conditions resulted in dual threonine and tyrosine phosphorylation of Slt2. Both Slt2 phosphorylation and FKS2-lacZ induction could be largely prevented by providing osmotic support to the plasma membrane. Interestingly, Slt2 phosphorylation in response to cell wall damage required the putative plasma-membrane-located sensor Mid2 but not Hcs77/Wsc1. Finally, cell wall perturbation gave rise to cells with increased resistance to glucanase digestion and heat shock. These responses depended on the presence of Slt2. These results indicate that weakening of the cell wall activates the Slt2/Mpk1 MAP kinase pathway and results in compensatory changes in the cell wall.
MeSH Terms
Benzenesulfonates/pharmacology
Calcium-Binding Proteins/metabolism
Cell Wall/drug effects,genetics,metabolism
Fluorescent Dyes/pharmacology
Fungal Proteins/metabolism
Glucan 1,3-beta-Glucosidase
Glucosyltransferases
Hot Temperature
Hydrolases/metabolism
Intracellular Signaling Peptides and Proteins
Lac Operon
Membrane Glycoproteins
Membrane Proteins/genetics,metabolism
Mitogen-Activated Protein Kinases
Phosphorylation
Recombinant Fusion Proteins/metabolism
Saccharomyces cerevisiae/genetics,physiology
Saccharomyces cerevisiae Proteins
Up-Regulation
beta-Glucosidase/metabolism
Chemicals
Benzenesulfonates
Calcium-Binding Proteins
Fluorescent Dyes
Fungal Proteins
Intracellular Signaling Peptides and Proteins
MID2 protein, S cerevisiae
Membrane Glycoproteins
Membrane Proteins
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
zymolyase
C.I. Fluorescent Brightening Agent 28
Glucosyltransferases
GSC2 protein, S cerevisiae
Mitogen-Activated Protein Kinases
SLT2 protein, S cerevisiae
Hydrolases
beta-Glucosidase
Glucan 1,3-beta-Glucosidase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
de Nobel Hans
Center for Fungal Cell Wall Research, Swammerdam Institute for Life Sciences, BioCentrum, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands1.
Ruiz Cristina
Departamento de Microbiologı́a II, Facultad Farmacia, UCM, Avda. Ramón y Cajal s/n 28040-Madrid, Spain2.
Martin Humberto
Departamento de Microbiologı́a II, Facultad Farmacia, UCM, Avda. Ramón y Cajal s/n 28040-Madrid, Spain2.
Morris Wayne
Department of Microbiology and Preservation, Unilever Research Laboratory, Olivier van Noortlaan 120, 3133 AT Vlaardingen, The Netherlands3. | Center for Fungal Cell Wall Research, Swammerdam Institute for Life Sciences, BioCentrum, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands1.
Brul Stanley
Department of Microbiology and Preservation, Unilever Research Laboratory, Olivier van Noortlaan 120, 3133 AT Vlaardingen, The Netherlands3. | Center for Fungal Cell Wall Research, Swammerdam Institute for Life Sciences, BioCentrum, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands1.
Molina Maria
Departamento de Microbiologı́a II, Facultad Farmacia, UCM, Avda. Ramón y Cajal s/n 28040-Madrid, Spain2.
Klis Frans M
Center for Fungal Cell Wall Research, Swammerdam Institute for Life Sciences, BioCentrum, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands1.