Functional interplay between Mediator and TFIIB in preinitiation complex assembly in relation to promoter architecture.
Mediator is a large coregulator complex conserved from yeast to humans and involved in many human diseases, including cancers. Together with general transcription factors, it stimulates preinitiation complex (PIC) formation and activates RNA polymerase II (Pol II) transcription. In this study, we analyzed how Mediator acts in PIC assembly using in vivo, in vitro, and in silico approaches. We revealed an essential function of the Mediator middle module exerted through its Med10 subunit, implicating a key interaction between Mediator and TFIIB. We showed that this Mediator-TFIIB link has a global role on PIC assembly genome-wide. Moreover, the amplitude of Mediator's effect on PIC formation is gene-dependent and is related to the promoter architecture in terms of TATA elements, nucleosome occupancy, and dynamics. This study thus provides mechanistic insights into the coordinated function of Mediator and TFIIB in PIC assembly in different chromatin contexts.
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Thomas
T
Eychenne
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Elizaveta
E
Novikova
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Marie-Bénédicte
MB
Barrault
Laboratoire d'Exploration Fonctionnelle des Génomes (LEFG), Institut de Radiobiologie Cellulaire et Moléculaire (IRCM), CEA, Genopole G2, F-91057 Evry Cedex, France.
Olivier
O
Alibert
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Claire
C
Boschiero
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Nuno
N
Peixeiro
Service d'Identification et de Caractérisation des Protéines par Spectrométrie de Masse (SICaPS), CNRS, F-91198 Gif-sur-Yvette Cedex, France.
David
D
Cornu
Service d'Identification et de Caractérisation des Protéines par Spectrométrie de Masse (SICaPS), CNRS, F-91198 Gif-sur-Yvette Cedex, France.|Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS, F-91198 Gif-sur-Yvette Cedex, France.
Virginie
V
Redeker
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.
Laurent
L
Kuras
Mathematiques et Informatique Appliquées du Génome à l'Environnement (MaIAGE), Institut National de la Recherche Agronomique (INRA), Université Paris-Saclay, 78350 Jouy-en-Josas, France.
Pierre
P
Nicolas
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Michel
M
Werner
Institute for Integrative Biology of the Cell (I2BC), Commissariat à l'Énergie Atomique (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris Sud, Université Paris Saclay, F-91198 Gif-sur-Yvette Cedex, France.|Institut de Biologie et de Technologies de Saclay (IBITECS), CEA, F-91191 Gif-sur-Yvette Cedex, France.
Julie
J
Soutourina
D002843:Chromatin / Q000378:metabolism; D056892:Mediator Complex / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011485:Protein Binding / Q000235:genetics; D055503:Protein Multimerization / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D035581:Transcription Factor TFIIB / Q000378:metabolism*
D002843:Chromatin; D056892:Mediator Complex; D029701:Saccharomyces cerevisiae Proteins; D035581:Transcription Factor TFIIB
Mediator; RNA polymerase II transcription; Saccharomyces cerevisiae; TFIIB; preinitiation complex; promoter architecture
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