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Rsc4 Connects the Chromatin Remodeler RSC to RNA Polymerases

Julie Soutourina,^1, Véronique Bordas-Le Floch,^1, Gabrielle Gendrel,^1, Amando Flores,^1, Cécile Ducrot,¹ Hélène Dumay-Odelot,^1,|| Pascal Soularue,² Francisco Navarro,³ Bradley R. Cairns,⁴ Olivier Lefebvre,¹ and Michel Werner^1*

Service de Biochimie et Génétique Moléculaire, Bâtiment 144, CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France,¹ Service de Génomique Fonctionnelle, CEA/Evry, 2 Rue Gaston Crémieux, CP22 F-91057 Evry Cedex, France,² Department Biología Experimental-Area de Genética (ED.B3), Universidad de Jaén, Parage la Lagunillas, E-23071 Jaén, Spain,³ Howard Hughes Medical Institute and Department of Oncological Science, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah 84112⁴

Received 9 March 2006/ Returned for modification 6 April 2006/ Accepted 13 April 2006

RSC is an essential, multisubunit chromatin remodeling complex. We show here that the Rsc4 subunit of RSC interacted via its C terminus with Rpb5, a conserved subunit shared by all three nuclear RNA polymerases (Pol). Furthermore, the RSC complex coimmunoprecipitated with all three RNA polymerases. Mutations in the C terminus of Rsc4 conferred a thermosensitive phenotype and the loss of interaction with Rpb5. Certain thermosensitive rpb5 mutations were lethal in combination with an rsc4 mutation, supporting the physiological significance of the interaction. Pol II transcription of ca. 12% of the yeast genome was increased or decreased twofold or more in a rsc4 C-terminal mutant. The transcription of the Pol III-transcribed genes SNR6 and RPR1 was also reduced, in agreement with the observed localization of RSC near many class III genes. Rsc4 C-terminal mutations did not alter the stability or assembly of the RSC complex, suggesting an impact on Rsc4 function. Strikingly, a C-terminal mutation of Rsc4 did not impair RSC recruitment to the RSC-responsive genes DUT1 and SMX3 but rather changed the chromatin accessibility of DNases to their promoter regions, suggesting that the altered transcription of DUT1 and SMX3 was the consequence of altered chromatin remodeling.

Supplemental material for this article may be found at http://mcb.asm.org/.

J.S., V.B.-L.F., and G.G. contributed equally to this study.

Present address: Departamento de Biología Molecular e Ingeniería Bioquímica, Centro Andaluz de Biología del Desarollo, Universidad Pablo de Olavide, Ctra. Utrera Km 1, E-41013 Sevilla, Spain.

^|| Present address: Institut Européen de Chimie et Biologie, Université Victor Segalen Bordeaux 2, 2, Rue Robert Escarpit, F-33607 Pessac, France.

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