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Eukaryotic Cell, March 2006, p. 507-517, Vol. 5, No. 3
1535-9778/06/$08.00+0     doi:10.1128/EC.5.3.507-517.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The RIM101 Pathway Contributes to Yeast Cell Wall Assembly and Its Function Becomes Essential in the Absence of Mitogen-Activated Protein Kinase Slt2p

F. Castrejon, A. Gomez, M. Sanz, A. Duran, and C. Roncero*

Instituto de Microbiologia Bioquimica and Departamento de Microbiologia y Genetica, CSIC/Universidad Salamanca, Salamanca, Spain

Received 7 December 2005/ Accepted 27 December 2005

The Saccharomyces cerevisiae ynl294c{Delta} (rim21{Delta}) mutant was identified in our lab owing to its moderate resistance to calcofluor, although it also displayed all of the phenotypic traits associated with its function as the putative sensor (Rim21p) of the RIM101 pathway. rim21{Delta} also showed moderate hypersensitivity to sodium dodecyl sulfate, caffeine, and zymolyase, and the cell wall compensatory response in this mutant was very poor, as indicated by the almost complete absence of Slt2 phosphorylation and the modest increase in chitin synthesis after calcofluor treatment. However, the cell integrity pathway appeared functional after caffeine treatment or thermal stress. rim21{Delta} and rim101{Delta} mutant strains shared all of the cell-wall-associated phenotypes, which were reverted by the expression of Rim101-531p, the constitutively active form of this transcription factor. Therefore, the absence of a functional RIM101 pathway leads to cell wall defects. rim21{Delta}, as well as rim101{Delta}, was synthetic lethal with slt2{Delta}, a synthetic defect alleviated by osmotic stabilization of the media. The double mutants grown in osmotically stabilized media were extremely hypersensitive to zymolyase and showed thicker cell walls, with poorly defined mannoprotein layers. In contrast, rim21{Delta} rlm1{Delta} and rim101{Delta} rlm1{Delta} double mutants were fully viable. Taken together, these results show that the RIM101 pathway participates directly in cell wall assembly and that it acts in parallel with the protein kinase C pathway (PKC) in this process independently of the transcriptional effect of the compensatory response mediated by this route. In addition, these results provide new experimental evidence of the direct involvement of the PKC signal transduction pathway through the Sltp2 kinase in the construction of yeast cell walls.

* Corresponding author. Mailing address: Departamento de Microbiologia y Genetica, CSIC/Universidad de Salamanca, Edificio Departamental, R-219, Avda. Campo Charro s/n, 37007-Salamanca, Spain. Phone: 34-923-294733. Fax: 34-923-224876. E-mail: crm{at}usal.es.

Eukaryotic Cell, March 2006, p. 507-517, Vol. 5, No. 3
1535-9778/06/$08.00+0     doi:10.1128/EC.5.3.507-517.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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