Journal article
Sedimentation and Immunoprecipitation Assays for Analyzing Complexes that Repress Transcription
Methods in molecular biology (Clifton, N.J.), Vol.977, pp.365-383
2013
DOI: 10.1007/978-1-62703-284-1_30
PMCID: PMC3653426
PMID: 23436378
Abstract
Co-repressor proteins function as platforms for the assembly of multi-subunit complexes that mediate transcriptional repression. Common components of such complexes are histone deacetylases, which catalyze the removal of acetyl groups from the tails of histones within nucleosomes, resulting in chromatin compaction and gene repression. In addition, co-repressor complexes generally interact with sequence-specific DNA-binding proteins that direct association with regulatory elements in the genome. Thus, identifying proteins that stably associate with co-repressors can provide insights regarding the biochemical function and target gene specificity of these molecules. Here, we describe a density gradient fractionation method for determining whether a co-repressor is incorporated into high-molecular complexes within cells and for identifying potential constituents of these complexes. We also describe a co-immunoprecipitation assay for confirming and further studying interactions between co-repressors and other proteins that may represent functional partners.
Details
- Title: Subtitle
- Sedimentation and Immunoprecipitation Assays for Analyzing Complexes that Repress Transcription
- Creators
- Ping Lu - Interdisciplinary Graduate Program in Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa Iowa, City, IA 52242Bruce S Hostager - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa Iowa, City, IA 52242Paul B Rothman - Interdisciplinary Graduate Program in Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa Iowa, City, IA 52242John D Colgan - Interdisciplinary Graduate Program in Immunology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa Iowa, City, IA 52242
- Resource Type
- Journal article
- Publication Details
- Methods in molecular biology (Clifton, N.J.), Vol.977, pp.365-383
- DOI
- 10.1007/978-1-62703-284-1_30
- PMID
- 23436378
- PMCID
- PMC3653426
- NLM abbreviation
- Methods Mol Biol
- eISBN
- 1627032843; 9781627032841
- ISSN
- 1064-3745
- eISSN
- 1940-6029
- Grant note
- R01 AI093737 || AI / National Institute of Allergy and Infectious Diseases Extramural Activities : NIAID R56 AI093737 || AI / National Institute of Allergy and Infectious Diseases Extramural Activities : NIAID
- Language
- English
- Date published
- 2013
- Academic Unit
- Microbiology and Immunology; Anatomy and Cell Biology; Immunology; Internal Medicine
- Record Identifier
- 9984025591002771
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