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Related Experiment Videos

A mechanism for coordinating chromatin modification and preinitiation complex assembly.

Joshua C Black1, Janet E Choi, Sarah R Lombardo

  • 1Department of Biological Chemistry, David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA.

Molecular Cell
|September 16, 2006
PubMed
Summary
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Mediator protein complex coordinates gene transcription by regulating histone acetyltransferases (HATs) and general transcription factors (GTFs). Autoacetylation of p300 HAT causes its dissociation, enabling GTF binding and transcription initiation.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Chromatin Dynamics

Background:

  • Eukaryotic gene transcription requires sequential recruitment of histone modifiers and general transcription factors (GTFs) by activators.
  • The coordination between chromatin modification and preinitiation complex assembly remains poorly understood.

Purpose of the Study:

  • To investigate how activators like GAL4-VP16 direct the ordered assembly of transcription machinery.
  • To elucidate the role of Mediator and histone acetyltransferases (HATs) in coordinating chromatin modification and transcription initiation.

Main Methods:

  • In vitro reconstitution assays using purified proteins and immobilized chromatin/DNA templates.
  • Investigated the interactions between Mediator, p300 (a HAT), and TFIID (a GTF).

Related Experiment Videos

  • Analyzed the effect of p300 autoacetylation on its catalytic activity, dissociation, and subsequent transcription factor binding.

    • Main Results:

    • GAL4-VP16 directed the ordered assembly of Mediator, HATs, and GTFs.
    • Mediator binds to both p300 and TFIID, regulating their assembly.
    • p300 autoacetylation induces a conformational change, leading to its dissociation from chromatin after acetylation.
    • p300 dissociation enhances TFIID binding and facilitates active transcription.

    Conclusions:

    • Autoacetylation-induced dissociation of p300 acts as a catalytic switch in transcription.
    • This switch allows for the timely binding of TFIID and the subsequent assembly of the preinitiation complex.
    • The Mediator complex plays a crucial role in coordinating these events for efficient gene transcription.