Characterization of nucleophosmin (B23) as a Myc target by scanning chromatin immunoprecipitation

J Biol Chem. 2001 Dec 21;276(51):48285-91. doi: 10.1074/jbc.M108506200. Epub 2001 Oct 16.

Abstract

The genetic program through which a specific transcription factor regulates a biological response is fundamental to our understanding how instructions in the genome are implemented. The emergence of DNA microarray technology for gene expression analysis has generated vast numbers of target genes resulting from specific transcription factor activity. We use the oncogenic transcription factor c-Myc as proof-of-principle that human genome sequence analysis and scanning of a specific gene by chromatin immunoprecipitation can be coupled to identify target transcription factor binding sequences. We focused on nucleophosmin, also known as B23, which was identified as a candidate Myc-responsive gene from a subtractive hybridization screen, and we found that sequences in intron 1, and not 5' sequences in the proximal promoter, are bound by c-Myc in vivo. Hence, a scanning chromatin immunoprecipitation (SChIP) strategy is useful in analyzing functional transcription factor-binding sites.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Animals
  • Base Sequence
  • Chromatin / metabolism*
  • DNA Primers
  • Gene Expression Profiling
  • Mice
  • Nuclear Proteins / metabolism*
  • Nucleic Acid Hybridization
  • Nucleophosmin
  • Polymerase Chain Reaction
  • Precipitin Tests
  • Protein Binding
  • Proto-Oncogene Proteins c-myc / metabolism*
  • Rats

Substances

  • Chromatin
  • DNA Primers
  • NPM1 protein, human
  • Npm1 protein, mouse
  • Nuclear Proteins
  • Proto-Oncogene Proteins c-myc
  • Nucleophosmin