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The Ccr4-Not Complex Regulates Skn7 through Srb10 Kinase

Eve Lenssen, Nowel Azzouz, Agnès Michel, Emilie Landrieux, and Martine A. Collart^*

Department of Microbiology and Molecular Medicine, University of Geneva Medical School, CMU, 1 rue Michel Servet, 1211 Geneva 4, Switzerland

Received 12 October 2006/ Accepted 9 October 2007

The Ccr4-Not complex is a multifunctional regulatory platform composed of nine subunits that controls diverse cellular events including mRNA degradation, protein ubiquitination, and transcription. In this study, we identified the yeast Saccharomyces cerevisiae osmotic and oxidative stress transcription factor Skn7 as a new target for regulation by the Ccr4-Not complex. Skn7 interacts with Not1 in a two-hybrid assay and coimmunoprecipitates with Not5 in a Not4-dependent manner. Skn7-dependent expression of OCH1 and Skn7 binding to the OCH1 promoter are increased in not4 or not5 mutants. Skn7 purified from wild-type cells but not from not4 cells is associated with the Srb10 kinase. This kinase plays a central role in the regulation of Skn7 by Not4, since increased OCH1 expression in not4 cells requires Srb10. These results reveal a critical role for the Ccr4-Not complex in the mechanism of activation of Skn7 that is dependent upon the Srb10 kinase.

Published ahead of print on 26 October 2007.

Present address: University of California Berkeley, 16 Barker Hall, MC 3204, Berkeley, CA 94720-3204.