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The transactivator proteins VP16 and GAL4 bind replication factor A

Z He1, B T Brinton, J Greenblatt

  • 1Department of Molecular and Medical Genetics, University of Toronto, Canada.

Cell
|June 18, 1993
PubMed

Insights

Transcription factors like VP16, GAL4, and p53 directly bind to replication factor A (RPA), a key protein in DNA replication initiation. This interaction is crucial for activating DNA replication and may involve stabilizing DNA or recruiting polymerase alpha.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Transcription factors play a role in initiating DNA replication.
  • Replication Factor A (RPA) is essential for DNA replication.
  • The interaction between transcription factors and RPA was previously unclear.

Purpose of the Study:

  • To investigate the direct interaction between transcription factors and RPA.
  • To identify the specific binding sites and subunits involved.
  • To understand the functional implications of this interaction in DNA replication.

Main Methods:

  • Affinity chromatography was used to purify and identify binding partners.
  • Specific transcription factors (VP16, GAL4, p53) and their acidic activation domains were tested.
  • Mutational analysis of VP16 was performed to assess binding and functional consequences.

Main Results:

  • The acidic activation domains of VP16, GAL4, and p53 selectively bind to human and yeast RPA.
  • This binding is direct and occurs with the largest subunit, RPA-1.
  • Mutations affecting VP16's ability to activate DNA replication also impaired its binding to RPA.

Conclusions:

  • Transcription factors directly interact with RPA-1, a subunit of RPA.
  • This interaction likely contributes to stabilizing single-stranded DNA at replication origins.
  • Transcription factors may recruit DNA polymerase alpha to the replication initiation complex via RPA.

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