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Related Experiment Videos

DA-complex assembly activity required for VP16C transcriptional activation

N Kobayashi1, P J Horn, S M Sullivan

  • 1Department of Microbiology and Molecular Genetics, Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095-1570, USA.

Molecular and Cellular Biology
|June 25, 1998
PubMed
Summary

The VP16C activation subdomain

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biochemistry

Background:

  • Transcriptional activation domains are crucial for gene expression.
  • Some domains facilitate the assembly of transcription factor IIA (TFIIA) and transcription factor IID (TFIID) complex on DNA.
  • The in vivo significance of this complex assembly activity remains to be fully elucidated.

Purpose of the Study:

  • To investigate the contribution of DA-complex assembly activity to the in vivo transcriptional activation mechanism.
  • To analyze the functional correlation between in vivo activation and in vitro DA-complex assembly using VP16C mutants.

Main Methods:

  • Site-directed mutagenesis of the VP16C activation subdomain.
  • In vivo transcriptional activation assays.

Related Experiment Videos

  • In vitro DNA-binding assays for interactions with TFIIA, TFIID, TFIIB, and TAFII40.

    • Main Results:

    • A strong correlation was found between in vivo activation function and in vitro DA-complex assembly activity for VP16C mutants.
    • Mutants deficient in in vivo activation exhibited reduced in vitro binding to TFIIA.
    • No significant correlation was observed between in vivo function and binding to TFIID subunits (TBP, TFIIB, TAFII40).

    Conclusions:

    • The DA-complex assembly activity of VP16C is a significant contributor to transcriptional activation in vivo.
    • VP16C's ability to enhance TFIIA and TFIID binding is critical for its activation function.