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NC2 mobilizes TBP on core promoter TATA boxes
Peter Schluesche1, Gertraud Stelzer, Elisa Piaia
1Department of Chemistry and Biochemistry and Center for Nanoscience (CeNS), Ludwig Maximilian University, Butenandtstr. 11, 81377 Munich, Germany.
Nature Structural & Molecular Biology
|November 13, 2007
Summary
Negative cofactor-2 (NC2) alters TATA-binding protein (TBP) dynamics on DNA. This finding challenges the assumption of direct GTF promoter recognition and suggests TBP can move away from the TATA box.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biophysics
Background:
- General transcription factors (GTFs) orchestrate eukaryotic RNA polymerase II transcription initiation.
- GTFs are believed to directly recognize promoter elements, forming stable preinitiation complexes.
- The TATA box-binding protein (TBP) specifically binds the TATA box, a key promoter element.
Purpose of the Study:
- To investigate the dynamic interactions between TBP, DNA, and regulatory factors during transcription initiation.
- To challenge the prevailing model of direct GTF promoter recognition.
- To elucidate the role of negative cofactor-2 (NC2) in modulating TBP-DNA complex stability.
Main Methods:
- Biochemical analysis of protein-DNA interactions.
- Biophysical single-pair Förster resonance energy transfer (spFRET) to monitor molecular dynamics.
- Investigating the effects of NC2 on TBP-DNA complex conformation.
Main Results:
- NC2 induces significant dynamic conformational changes in the TBP-DNA complex.
- These conformational changes enable TBP to transiently escape TATA-binding mode.
- Evidence suggests TBP can move along the DNA away from the TATA box.
- TBP can re-bind to the TATA box after escaping.
Conclusions:
- NC2 plays a crucial role in regulating TBP dynamics at the promoter.
- The TBP-TATA box interaction is not static, allowing for promoter escape and repositioning.
- This dynamic mechanism may influence transcription initiation site selection and regulation.
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