Chapter title |
Assessing Sites of NF-κB DNA Binding Using Chromatin Immunoprecipitation.
|
---|---|
Chapter number | 4 |
Book title |
NF-kappa B
|
Published in |
Methods in molecular biology, February 2015
|
DOI | 10.1007/978-1-4939-2422-6_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2421-9, 978-1-4939-2422-6
|
Authors |
Amy Colleran, Patricia E Collins, Ruaidhrí J Carmody, Patricia E. Collins, Ruaidhrí J. Carmody, Colleran, Amy, Collins, Patricia E., Carmody, Ruaidhrí J. |
Editors |
Michael J. May |
Abstract |
The NF-κB transcription factor is in fact a family of related proteins which dimerize to form at least 12 distinct complexes which regulate the expression of hundred of genes of importance to a range of physiological and pathological processes. The binding of NF-κB to the regulatory regions and promoters of target genes is influenced by a number of factors including the sequence of DNA-binding sites, the posttranslational modification of NF-κB, and the interaction of cofactors and co-regulators of transcription. In addition, the binding of NF-κB to promoters is highly dynamic and the recruitment of specific subunits to specific binding sites may occur with distinct kinetics. Moreover, genome-wide analysis of NF-κB chromatin binding indicates that the majority of DNA-binding events are not associated with changes in transcriptional activity. Thus, the analysis of NF-κB recruitment and activity at specific binding sites is of critical importance in understanding the regulation of transcription. In this chapter we describe a chromatin immunoprecipitation assay to investigate the in situ binding of NF-κB to specific sites in the genome. |
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