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Identification of a Myb-responsive enhancer of the chicken C/EBPbeta gene

Jörg Kintscher1, Vichanan Yamkamon, Daniel Braas

  • 1Institut für Biochemie, Westfälische-Wilhelms-Universität Münster, Wilhelm-Klemm-Str. 2, D-48149 Münster, Germany.

Oncogene
|June 15, 2004
PubMed

Insights

The retroviral oncogene v-Myb disrupts cell differentiation by altering gene expression. This study identifies a novel enhancer element regulated by v-Myb, revealing new insights into its mechanism of action in myelomonocytic cells.

Area of Science:

  • * Molecular Biology
  • * Cancer Biology
  • * Gene Regulation

Background:

  • * The retroviral oncogene v-myb encodes v-Myb, a transcription factor that disrupts myelomonocytic differentiation and transforms cells.
  • * v-Myb's biological effects are attributed to the deregulation of specific target genes, including CCAAT box/enhancer binding protein beta (C/EBPbeta).
  • * C/EBPbeta is crucial for myelomonocytic differentiation and is known to be regulated by Myb.

Purpose of the Study:

  • * To elucidate the mechanism by which v-Myb affects C/EBPbeta expression.
  • * To identify and characterize v-Myb target sites in vivo using chromatin structure analysis.
  • * To investigate the role of a novel downstream enhancer element in C/EBPbeta gene regulation by v-Myb.

Main Methods:

  • * Mapping of DNase I hypersensitive sites (DHSs) in chromatin to detect in vivo v-Myb target sites.
  • * Chromatin immunoprecipitation (ChIP) to confirm v-Myb binding to identified DHSs.
  • * Reporter gene assays in transiently and stably transfected myelomonocytic cells to assess enhancer activity.

Main Results:

  • * Identification of a novel DHS downstream of the C/EBPbeta gene specific to v-myb-transformed myeloblasts.
  • * Confirmation of v-Myb binding to this downstream region in vivo.
  • * Demonstration that v-Myb induces this DHS, indicating an effect on chromatin structure.
  • * Characterization of the downstream DHS as a Myb-dependent enhancer element.
  • * Discovery that v-Myb regulates C/EBPbeta via both promoter and enhancer elements, with distinct mechanisms.
  • * Promoter activation requires C/EBPbeta cooperation, while enhancer activation is independent of C/EBPbeta.

Conclusions:

  • * v-Myb stimulates C/EBPbeta expression through both promoter and enhancer elements.
  • * A novel Myb-dependent enhancer downstream of the C/EBPbeta gene has been identified.
  • * v-Myb directly alters chromatin structure to regulate gene expression.
  • * Chromatin structure analysis is a valuable tool for identifying transcription factor target sites.

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