title: The nucleosomal barrier to promoter escape by RNA polymerase II is overcome by the chromatin remodeler Chd1.
abstract: RNA polymerase II (PolII) transcribes RNA within a chromatin context, with nucleosomes acting as barriers to transcription. Despite these barriers, transcription through chromatin in vivo is highly efficient, suggesting the existence of factors that overcome this obstacle. To increase the resolution obtained by standard chromatin immunoprecipitation, we developed a novel strategy using micrococcal nuclease digestion of cross-linked chromatin. We find that the chromatin remodeler Chd1 is recruited to promoter proximal nucleosomes of genes undergoing active transcription, where Chd1 is responsible for the vast majority of PolII-directed nucleosome turnover. The expression of a dominant negative form of Chd1 results in increased stalling of PolII past the entry site of the promoter proximal nucleosomes. We find that Chd1 evicts nucleosomes downstream of the promoter in order to overcome the nucleosomal barrier and enable PolII promoter escape, thus providing mechanistic insight into the role of Chd1 in transcription and pluripotency. DOI: http://dx.doi.org/10.7554/eLife.02042.001.
authors.affiliation: Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Skene
authors.forename: Aaron E
authors.initials: AE
authors.lastname: Hernandez
authors.forename: Mark
authors.initials: M
authors.lastname: Groudine
authors.forename: Steven
authors.initials: S
authors.lastname: Henikoff
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: C489844:Chd1 protein, mouse; D004268:DNA-Binding Proteins; D009707:Nucleosomes; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases
keywords: RNA polymerase II; chromatin; promoter escape; transcription
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