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J. Biol. Chem., Vol. 280, Issue 10, 9149-9159, March 11, 2005

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Pkc1 and the Upstream Elements of the Cell Integrity Pathway in Saccharomyces cerevisiae, Rom2 and Mtl1, Are Required for Cellular Responses to Oxidative Stress^*

Felipe Vilella, Enrique Herrero, Jordi Torres, and Maria Angeles de la Torre-Ruiz

From the Departament de Ciències Mèdiques Bàsiques, Alcalde Rovira Roure 44, Universitat de Lleida, Lleida 25198, Spain

In this study we analyze the participation of the PKC1-MAPK cell integrity pathway in cellular responses to oxidative stress in Saccharomyces cerevisiae. Evidence is presented demonstrating that only Pkc1 and the upstream elements of the cell integrity pathway are essential for cell survival upon treatment with two oxidizing agents, diamide and hydrogen peroxide. Mtl1 is characterized for the first time as a cell-wall sensor of oxidative stress. We also show that the actin cytoskeleton is a cellular target for oxidative stress. Both diamide and hydrogen peroxide provoke a marked depolarization of the actin cytoskeleton, being Mtl1, Rom2 and Pkc1 functions all required to restore the correct actin organization. Diamide induces the formation of disulfide bonds in newly secreted cell-wall proteins. This mainly provokes structural changes in the cell outer layer, which activate the PKC1-MAPK pathway and hence the protein kinase Slt2. Our results led us to the conclusion that Pkc1 activity is required to overcome the effects of oxidative stress by: (i) enhancing the machinery required to repair the altered cell wall and (ii) restoring actin cytoskeleton polarity by promoting actin cable formation.

Received for publication, September 27, 2004 , and in revised form, January 4, 2005.

^* This work was supported by the Spanish Ministry of Science and Technology (Grant BMC2001-1213-C02-01), the Instituto de Salud Carlos III (Spanish Ministry of Health and Consumption, Grant PI030734), and the Generalitat de Catalunya (Grant 2001-SGR-00305). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 34-973-702-409; Fax: 34-973-702-426; E-mail: madelatorre{at}cmb.udl.es.

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