Proteomic Analysis of Coregulators Bound to ERα on DNA and Nucleosomes Reveals Coregulator Dynamics
| Autor: | Chunshu Li, Jun Qin, Ross A. Hamilton, Sophia Y. Tsai, Zheng Zhang, Ming-Jer Tsai, Suzanna L. Bailey, Dean P. Edwards, Bert W. O'Malley, Doug W. Chan, Leah A. Gates, Amol O. Bajaj, Anna Malovannaya, Qin Feng, Charles E. Foulds, Chen Ding, David M. Lonard, Tamra L. Hunsaker, Celetta Callaway |
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| Rok vydání: | 2013 |
| Předmět: |
Proteomics
Transcriptional Activation Chromatin Immunoprecipitation Transcription Genetic Sialoglycoproteins Breast Neoplasms DNA-Activated Protein Kinase Transcription coregulator Biology Response Elements DNA-binding protein Article Nuclear Receptor Coactivator 3 Humans Nucleosome Phosphorylation Promoter Regions Genetic Enhancer Transcription factor Molecular Biology Forkhead Box Protein O1 Estrogen Receptor alpha Nuclear Proteins Estrogens Forkhead Transcription Factors DNA Cell Biology Molecular biology Peptide Fragments Nucleosomes Cell biology DNA-Binding Proteins Gene Expression Regulation Neoplastic Multiprotein Complexes Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization MCF-7 Cells Trans-Activators Female Transcription factor II D E1A-Associated p300 Protein Chromatin immunoprecipitation Estrogen receptor alpha hormones hormone substitutes and hormone antagonists HeLa Cells |
| Zdroj: | Molecular Cell. 51(2):185-199 |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2013.06.007 |
| Popis: | Chromatin immunoprecipitation studies have mapped protein occupancies at many genomic loci. However, a detailed picture of the complexity of coregulators (CoRs) bound to a defined enhancer along with a transcription factor is missing. To address this, we used biotin-DNA pull-down assays coupled with mass spectrometry-immunoblotting to identify at least 17 CoRs from nuclear extracts bound to 17β-estradiol (E2)-liganded estrogen receptor-α on estrogen response elements (EREs). Unexpectedly, these complexes initially are biochemically stable and contain certain atypical corepressors. Addition of ATP dynamically converts these complexes to an "activated" state by phosphorylation events, primarily mediated by DNA-dependent protein kinase. Importantly, a "natural" ERE-containing enhancer and nucleosomal EREs recruit similar complexes. We further discovered the mechanism whereby H3K4me3 stimulates ERα-mediated transcription as compared with unmodified nucleosomes. H3K4me3 templates promote specific CoR dynamics in the presence of ATP and AcCoA, as manifested by CBP/p300 and SRC-3 dismissal and SAGA and TFIID stabilization/recruitment. |
| Databáze: | OpenAIRE |
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