Characterizing the DNA binding site specificity of NF-κB with protein-binding microarrays (PBMs)

Methods Mol Biol. 2015:1280:609-30. doi: 10.1007/978-1-4939-2422-6_36.

Abstract

NF-κB transcription factors control a wide array of important cellular and organismal processes in eukaryotes. All NF-κB transcription factors bind to DNA target sites as dimers. In vertebrates, there are five NF-κB subunits, p50, p52, RelA (p65), c-Rel, and RelB, that can form almost all combinations of homodimers and heterodimers, which recognize distinct, but overlapping, target sequences. In this chapter, we describe the use of protein-binding microarrays (PBMs), a high-throughput method to measure the binding of proteins to different DNA sequences. PBM datasets allow for sensitive comparisons of NF-κB dimer DNA-binding differences and can aid in the computational and experimental prediction of NF-κB target genes.

Publication types

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

MeSH terms

  • Binding Sites*
  • DNA / genetics*
  • DNA / metabolism*
  • NF-kappa B / metabolism*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Protein Binding

Substances

  • NF-kappa B
  • DNA