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Functional interaction of adenovirus E1A with holo-TFIID
1Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024-1570.
Genes & Development
|September 1, 1993
Summary
Regulatory transcription factors bind TATA box-binding protein (TBP). Adenovirus E1A protein binds holo-TFIID, revealing TBP
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Transcription factors regulate gene expression by interacting with DNA and proteins.
- TATA box-binding protein (TBP) is a core transcription factor.
- TBP interacts with TBP-associated factors (TAFs) to form holo-TFIID, a complex crucial for transcription initiation.
Purpose of the Study:
- To investigate the interaction between the adenovirus E1A activation domain and the holo-TFIID complex.
- To determine if TBP-associated factors (TAFs) influence E1A binding to TBP.
- To identify the functional target of the E1A activation domain in transcription regulation.
Main Methods:
- In vitro binding assays to test E1A interaction with TBP and holo-TFIID.
- Establishment of an in vitro transcription system responsive to E1A.
- Squelching assays using excess E1A to identify the target factor.
Main Results:
- The E1A activation domain specifically binds to holo-TFIID, not just TBP alone.
- Excess E1A inhibits transcription by sequestering a required factor.
- Holo-TFIID is uniquely capable of reversing E1A-mediated transcriptional inhibition.
Conclusions:
- The E1A activation domain directly interacts with holo-TFIID.
- Holo-TFIID is the functional target of E1A in regulating transcription.
- This interaction is critical for E1A-dependent and E1A-independent transcription.