EP400 Deposits H3.3 into Promoters and Enhancers during Gene Activation

Suman K Pradhan; Trent Su; Linda Yen; Karine Jacquet; Chengyang Huang; Jacques Côté; Siavash K Kurdistani; Michael F Carey

doi:10.1016/j.molcel.2015.10.039

EP400 Deposits H3.3 into Promoters and Enhancers during Gene Activation

Mol Cell. 2016 Jan 7;61(1):27-38. doi: 10.1016/j.molcel.2015.10.039. Epub 2015 Dec 6.

Authors

Suman K Pradhan¹, Trent Su¹, Linda Yen², Karine Jacquet³, Chengyang Huang¹, Jacques Côté³, Siavash K Kurdistani⁴, Michael F Carey⁵

Affiliations

¹ Department of Biological Chemistry, David Geffen School of Medicine, UCLA, 351A Biomedical Sciences Research Building, 615 Charles E. Young Drive South, Los Angeles, CA 90095-1737, USA.
² The Molecular Biology Institute, UCLA, Paul D. Boyer Hall, 611 Charles E. Young Drive South, Los Angeles, CA 90095-1570, USA.
³ Laval University Cancer Research Center, CHU de Québec Research Center-Oncology, Hôtel-Dieu de Québec, 9 McMahon Street, Quebec City, QC G1R 2J6, Canada.
⁴ Department of Biological Chemistry, David Geffen School of Medicine, UCLA, 351A Biomedical Sciences Research Building, 615 Charles E. Young Drive South, Los Angeles, CA 90095-1737, USA; The Molecular Biology Institute, UCLA, Paul D. Boyer Hall, 611 Charles E. Young Drive South, Los Angeles, CA 90095-1570, USA.
⁵ Department of Biological Chemistry, David Geffen School of Medicine, UCLA, 351A Biomedical Sciences Research Building, 615 Charles E. Young Drive South, Los Angeles, CA 90095-1737, USA; The Molecular Biology Institute, UCLA, Paul D. Boyer Hall, 611 Charles E. Young Drive South, Los Angeles, CA 90095-1570, USA. Electronic address: mcarey@mednet.ucla.edu.

Abstract

Gene activation in metazoans is accompanied by the presence of histone variants H2AZ and H3.3 within promoters and enhancers. It is not known, however, what protein deposits H3.3 into chromatin or whether variant chromatin plays a direct role in gene activation. Here we show that chromatin containing acetylated H2AZ and H3.3 stimulates transcription in vitro. Analysis of the Pol II pre-initiation complex on immobilized chromatin templates revealed that the E1A binding protein p400 (EP400) was bound preferentially to and required for transcription stimulation by acetylated double-variant chromatin. EP400 also stimulated H2AZ/H3.3 deposition into promoters and enhancers and influenced transcription in vivo at a step downstream of the Mediator complex. EP400 efficiently exchanged recombinant histones H2A and H3.1 with H2AZ and H3.3, respectively, in a chromatin- and ATP-stimulated manner in vitro. Our data reveal that EP400 deposits H3.3 into chromatin alongside H2AZ and contributes to gene regulation after PIC assembly.

Keywords: EP400; H2AZ; H3.3; RNA polymerase II; histone exchange; pre-initiation complex (PIC).

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Acetylation
Adenosine Triphosphate / metabolism
Binding Sites
Cell Line, Tumor
Chromatin Assembly and Disassembly*
DNA Helicases / genetics
DNA Helicases / metabolism*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Gene Expression Regulation*
Genes, Reporter
Histones / genetics
Histones / metabolism*
Humans
Promoter Regions, Genetic*
RNA Interference
RNA Polymerase II / metabolism
Time Factors
Transcriptional Activation*
Transfection

Substances

DNA-Binding Proteins
Histones
histone H2A.F-Z
Adenosine Triphosphate
RNA Polymerase II
DNA Helicases
EP400 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding