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J. Biol. Chem., Vol. 282, Issue 23, 16838-16845, June 8, 2007

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Regulation of Snf1 Protein Kinase in Response to Environmental Stress^*

From the Departments of Genetics & Development and Microbiology, Columbia University, New York, New York 10032

The Saccharomyces cerevisiae Snf1 protein kinase, a member of the Snf1/AMPK (AMP-activated protein kinase) family, has important roles in metabolic control, particularly in response to nutrient stress. Here we have addressed the role of Snf1 in responses to other environmental stresses. Exposure of cells to sodium ion stress, alkaline pH, or oxidative stress caused an increase in Snf1 catalytic activity and phosphorylation of Thr-210 in the activation loop, whereas treatment with sorbitol or heat shock did not. Inhibition of respiratory metabolism by addition of antimycin A to cells also increased Snf1 activity. Analysis of mutants indicated that the kinases Sak1, Tos3, and Elm1, which activate Snf1 in response to glucose limitation, are also required under other stress conditions. Each kinase sufficed for activation in response to stress, but Sak1 had the major role. In sak1 tos3 elm1 cells expressing mammalian Ca²⁺/calmodulin-dependent protein kinase kinase , Snf1 was activated by both sodium ion and alkaline stress, suggesting that stress signals regulate Snf1 activity by a mechanism that is independent of the upstream kinase. Finally, we showed that Snf1 protein kinase is regulated differently during adaptation of cells to NaCl and alkaline pH with respect to both temporal regulation of activation and subcellular localization. Snf1 protein kinase becomes enriched in the nucleus in response to alkaline pH but not salt stress. Such differences could contribute to specificity of the stress responses.

Received for publication, January 5, 2007 , and in revised form, April 16, 2007.

^* This work was supported by National Institutes of Health Grant GM34095 (to M. C.). 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.

¹ To whom correspondence should be addressed: 701 W. 168th St., HSC922, New York, NY 10032. Tel.: 212-305-6314; Fax: 212-305-1741; E-mail: mbc1{at}columbia.edu.

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