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Related Experiment Videos

Multiple N-CoR complexes contain distinct histone deacetylases.

P L Jones1, L M Sachs, N Rouse

  • 1Unit of Molecular Morphogenesis, Laboratory of Molecular Embryology, NICHD, National Institutes of Health, Bethesda, Maryland 20892-5431 , USA.

The Journal of Biological Chemistry
|March 20, 2001
PubMed
Summary
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Nuclear receptor corepressor (N-CoR) forms novel complexes with histone deacetylases (HDACs) in Xenopus eggs. These N-CoR-HDAC complexes are involved in transcriptional repression by unliganded thyroid hormone receptors (TRs).

Area of Science:

  • Molecular Biology
  • Epigenetics
  • Transcriptional Regulation

Background:

  • Nuclear receptor corepressor (N-CoR) mediates transcriptional repression by unliganded nuclear receptors, including thyroid hormone receptors (TRs).
  • N-CoR is known to interact with Sin3 and histone deacetylase Rpd3 (HDAC1) in vitro, suggesting a corepressor complex.
  • Previous studies failed to biochemically isolate endogenous N-CoR-containing complexes with histone deacetylase (HDAC) activity.

Purpose of the Study:

  • To biochemically characterize N-CoR-containing complexes involved in transcriptional repression.
  • To investigate the association of N-CoR with histone deacetylases in Xenopus oocytes.
  • To determine if N-CoR complexes are recruited by unliganded TRs for gene silencing.

Main Methods:

Related Experiment Videos

  • Biochemical fractionation of Xenopus egg extracts to isolate macromolecular complexes.
  • Histone deacetylase activity assays to identify enzymatic complexes.
  • Immunoprecipitation studies to confirm protein-protein interactions in frog oocytes.

    • Main Results:

    • Three novel macromolecular complexes containing N-CoR were isolated from Xenopus egg extract.
    • Two of these complexes exhibited histone deacetylase activity: one dependent on Sin3 and Rpd3, and another Sin3-independent complex.
    • The third complex contained N-CoR but lacked detectable HDAC activity.
    • N-CoR was shown to bind unliganded TR in frog oocytes, confirming its role in TR-mediated repression.

    Conclusions:

    • N-CoR associates with distinct histone deacetylase complexes, both Sin3-dependent and Sin3-independent.
    • These N-CoR-HDAC complexes are biochemically isolated and functionally implicated in transcriptional repression by unliganded TRs.
    • N-CoR utilizes at least two different HDAC pathways to target specific gene promoters for repression.