Characterization of nucleophosmin (B23) as a Myc target by scanning chromatin immunoprecipitation.
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.
Zeller
K I
KI
Program in Human Genetics and Molecular Biology and Department of Medicine, The Johns Hopkins Oncology Center and The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Haggerty
T J
TJ
Barrett
J F
JF
Guo
Q
Q
Wonsey
D R
DR
Dang
C V
CV
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002843:Chromatin / Q000378:metabolism*; D017931:DNA Primers; D020869:Gene Expression Profiling; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D000090243:Nucleophosmin; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011485:Protein Binding; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D051381:Rats
D002843:Chromatin; D017931:DNA Primers; C000716967:NPM1 protein, human; C000715570:Npm1 protein, mouse; D009687:Nuclear Proteins; D016271:Proto-Oncogene Proteins c-myc; D000090243:Nucleophosmin