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The adenovirus E1A repression domain disrupts the interaction between the TATA binding protein and the TATA box in a

C Z Song1, P M Loewenstein, K Toth

  • 1Institute for Molecular Virology, Saint Louis University School of Medicine, Missouri 63110, USA.

Molecular and Cellular Biology
|April 1, 1997
PubMed

Insights

Human adenovirus E1A oncoprotein directly binds TATA-binding protein (TBP) to repress transcription. This interaction blocks TBP-TATA box binding, a process reversible by TFIIB, revealing a novel repression mechanism.

Area of Science:

  • Molecular Biology
  • Virology
  • Gene Regulation

Background:

  • Human adenovirus E1A oncoprotein has a transcription repression function linked to cell cycle control and differentiation inhibition.
  • The precise molecular mechanism of E1A-mediated transcriptional repression remains incompletely understood.
  • Previous work identified TATA-binding protein (TBP) as a cellular target of E1A repression.

Purpose of the Study:

  • To elucidate the molecular mechanism by which adenovirus E1A represses transcription.
  • To characterize the direct interaction between E1A and TBP.
  • To investigate the role of TFIIB in modulating E1A-mediated repression.

Main Methods:

  • Demonstration of direct and specific binding between TBP and the E1A repression domain.
  • Mapping of the TBP binding domain within E1A 243R to N-terminal residues 1-35.
  • Gel mobility shift and DNase protection assays using an E1A 1-80 fragment to assess TBP-TATA box interactions.
  • In vitro transcription assays to evaluate the effects of E1A and TFIIB.

Main Results:

  • The interaction between TBP and the E1A repression domain is direct and specific.
  • The E1A N-terminal residues 1-35 form the TBP binding domain, distinct from the conserved region 3 domain.
  • E1A 1-80 fragment inhibits TBP binding to the TATA box and can dissociate preformed TBP-TATA complexes.
  • TFIIB prevents E1A-mediated disruption of TBP-TATA interactions and can overcome E1A repression in vitro.

Conclusions:

  • Adenovirus E1A represses transcription by directly interfering with TBP binding to the TATA box.
  • TFIIB plays a crucial role in reversing E1A-mediated repression by stabilizing the TBP-TATA complex.
  • This mechanism of E1A repression is distinct from known cellular repressors targeting TBP.

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