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Related Experiment Videos

CHF: a novel factor binding to cyclin A CHR corepressor element.

A Philips1, S Chambeyron, N Lamb

  • 1Institut de Génétique Moléculaire, CNRS, UMR 5535, Montpellier, France.

Oncogene
|December 22, 1999
PubMed
Summary

Researchers identified a novel protein, cyclin A CHR binding factor (CHF), that represses cyclin A gene expression during the cell cycle. This discovery sheds light on the intricate regulation of cell proliferation and gene transcription.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Gene Regulation

Background:

  • Cyclin A expression is tightly regulated during the cell cycle through transcriptional repression.
  • This repression involves a bipartite negative DNA regulatory region: Cell Cycle Responsive Element/Cell Cycle Dependent Element (CCRE/CDE) and Cell Cycle gene Homology Region (CHR).
  • While E2F/DP binding to CCRE/CDE is suggested, the proteins interacting with CHR remain unidentified.

Purpose of the Study:

  • To identify and characterize proteins that bind to the cyclin A CHR.
  • To understand the functional role of CHR-binding proteins in cyclin A gene regulation.
  • To elucidate differences in repressor region function between cell cycle-regulated genes.

Main Methods:

  • Characterization of nuclear extracts from quiescent and S-phase blocked cells.

Related Experiment Videos

  • Electrophoretic mobility shift assays (EMSA) and UV crosslinking to identify CHR-binding proteins.
  • Oligonucleotide competition assays, in vitro transcription, and in vivo microinjection experiments.

    • Main Results:

    • An activity binding specifically to cyclin A CHR, but not to CHRs of other cell cycle genes, was identified.
    • A 90 kD protein, named cyclin A CHR binding factor (CHF), was identified.
    • Functional differences were observed between the repressor regions of cyclin A and cdc25C genes.

    Conclusions:

    • A novel 90 kD protein, CHF, specifically binds to the cyclin A CHR and is involved in transcriptional repression.
    • CHF binding activity is cell cycle-dependent, being present in quiescent cells and absent in S-phase blocked cells.
    • The study reveals distinct mechanisms of transcriptional repression mediated by composite regulatory regions in cell cycle-controlled genes.