A High-Throughput Chromatin Immunoprecipitation Sequencing Approach to Study the Role of MYC on the Epigenetic Landscape

Methods Mol Biol. 2021:2318:187-208. doi: 10.1007/978-1-0716-1476-1_9.

Abstract

MYC is a transcription factor playing multiple functions both in physiological and pathological settings. Biochemical characterizations, combined with the analyses of MYC chromatin binding, have shown that its pleiotropic activity depends on the chromatin context and its protein-protein interactions with different cofactors. In order to determine the contribution of MYC in a certain biological condition, it would be relevant to analyze the concomitant binding of MYC and its associated proteins, in relationship to the chromatin environment. To this end, we here provide a simple method to parallel map the genome-wide binding of MYC-associated proteins, together with the chromatin profiling of multiple histone modifications. We detail the procedure to perform high-throughput ChIP-seq (HT-ChIP-seq) with a variety of biological samples. In addition, we describe simple bioinformatic steps to determine the distribution of MYC binding with respect to the chromatin context and the association of its cofactors. The described approach will permit the reproducible characterization of MYC activity in different biological contexts.

Keywords: ChIP-seq; Chromatin; Epigenetics; Histone modifications; MYC.

MeSH terms

  • Chromatin / genetics
  • Chromatin Immunoprecipitation / methods
  • Chromatin Immunoprecipitation Sequencing / methods*
  • Computational Biology / methods
  • DNA / genetics
  • Epigenesis, Genetic / genetics
  • Epigenomics / methods*
  • Genes, myc / genetics
  • Genes, myc / physiology
  • High-Throughput Nucleotide Sequencing / methods
  • Histone Code / genetics
  • Histones / metabolism
  • Humans
  • Protein Processing, Post-Translational / genetics
  • Proto-Oncogene Proteins c-myc / genetics*
  • Proto-Oncogene Proteins c-myc / metabolism
  • Sequence Analysis, DNA / methods
  • Transcription Factors / metabolism

Substances

  • Chromatin
  • Histones
  • Proto-Oncogene Proteins c-myc
  • Transcription Factors
  • DNA