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c-Myb repression of c-erbB-2 transcription by direct binding to the c-erbB-2 promoter

G Mizuguchi1, C Kanei-Ishii, T Takahashi

  • 1Laboratory of Molecular Genetics, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.

The Journal of Biological Chemistry
|April 21, 1995
PubMed

Insights

The c-Myb protein represses the human c-erbB-2 gene promoter. This repression involves Myb binding sites, including one overlapping the TATA box, suggesting competition with TFIID for DNA binding.

Area of Science:

  • Molecular Biology
  • Oncogenes
  • Gene Regulation

Background:

  • The c-Myb proto-oncogene product (c-Myb) is a known transcriptional activator.
  • While c-Myb can repress artificial promoters, natural target genes repressed by c-Myb were unidentified.

Purpose of the Study:

  • To identify natural target genes transcriptionally repressed by c-Myb.
  • To elucidate the mechanism of c-Myb-mediated repression of the c-erbB-2 promoter.

Main Methods:

  • Chloramphenicol acetyltransferase (CAT) co-transfection assays were used to assess promoter activity.
  • Domain analyses of c-Myb were performed.
  • In vitro transcription assays and DNase I footprint analyses were conducted.

Main Results:

  • c-Myb and B-Myb were found to repress the human c-erbB-2 promoter activity.
  • Two specific Myb binding sites within the c-erbB-2 promoter were critical for repression.
  • c-Myb binding to a site overlapping the TATA box competed with TFIID binding.

Conclusions:

  • The human c-erbB-2 gene is a natural target gene transcriptionally repressed by c-Myb.
  • Myb-induced trans-repression of the c-erbB-2 promoter involves competition with TFIID for binding to the promoter region, particularly at the TATA box.

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