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The Ccr4-Not Complex Independently Controls both Msn2-Dependent Transcriptional Activation—via a Newly Identified Glc7/Bud14 Type I Protein Phosphatase Module—and TFIID Promoter Distribution

Eve Lenssen,^1, Nicole James,^1, Ivo Pedruzzi,^1, Frédérique Dubouloz,¹ Elisabetta Cameroni,¹ Ruth Bisig,¹ Laurent Maillet,^1, Michel Werner,² Johnny Roosen,³ Katarina Petrovic,^4, Joris Winderickx,³ Martine A. Collart,^1* and Claudio De Virgilio^1*

Departement de Microbiologie et Médecine Moléculaire, CMU, Geneva, and Botanisches Institut der Universität, Basel, Switzerland,¹ Service de Biochimie et de Génétique Moléculaire, CEA/Saclay, Gif-sur-Yvette, France,² Functional Biology, Katholieke Universiteit Leuven, Leuven-Heverlee, Flanders, Belgium³

Received 6 September 2004/ Accepted 29 September 2004

The Ccr4-Not complex is a conserved global regulator of gene expression, which serves as a regulatory platform that senses and/or transmits nutrient and stress signals to various downstream effectors. Presumed effectors of this complex in yeast are TFIID, a general transcription factor that associates with the core promoter, and Msn2, a key transcription factor that regulates expression of stress-responsive element (STRE)-controlled genes. Here we show that the constitutively high level of STRE-driven expression in ccr4-not mutants results from two independent effects. Accordingly, loss of Ccr4-Not function causes a dramatic Msn2-independent redistribution of TFIID on promoters with a particular bias for STRE-controlled over ribosomal protein gene promoters. In parallel, loss of Ccr4-Not complex function results in an alteration of the posttranslational modification status of Msn2, which depends on the type 1 protein phosphatase Glc7 and its newly identified subunit Bud14. Tests of epistasis as well as transcriptional analyses of Bud14-dependent transcription support a model in which the Ccr4-Not complex prevents activation of Msn2 via inhibition of the Bud14/Glc7 module in exponentially growing cells. Thus, increased activity of STRE genes in ccr4-not mutants may result from both altered general distribution of TFIID and unscheduled activation of Msn2.

E.L., N.J., and I.P. contributed equally to the present work.

Present address: Institut de Biochimie et Génétique Cellulaires, 33077 Bordeaux, France.

Present address: InPheno AG, CH-4051 Basel, Switzerland.

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