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M109246200v1
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Papers In Press, published online ahead of print October 24, 2001
J. Biol. Chem, 10.1074/jbc.M109246200
Submitted on September 25, 2001
Revised on October 24, 2001
Accepted on October 24, 2001

Fluorescence based analyses of the effects of full length recombinant TAF130p on the interaction of TBP with TATA box DNA

Utpal Banik, Joseph M Beechem, Edward Klebanow, Stephanie Schroeder, and Peter A Weil

Molecular Physiology & Biophysics, Vanderbilt University, School of Medicine, Nashville, TN 37232-0615

Corresponding Author: tony.weil{at}mcmail.vanderbilt.edu

We have used a combination of fluorescence anisotropy spectroscopy and fluorescence-based native gel electrophoresis methods to examine the effects of the TFIID-specific subunit TAF130p (TAF145p) upon the TATA-box DNA binding properties of TBP. Purified full length recombinant TAF130p decreases TBP-TATA DNA complex formation at equilibrium by competing directly with DNA for binding to TBP. Interestingly, we have found that full length TAF130p is capable of binding multiple molecules of TBP with nM binding affinity. The biological implications of these findings are discussed.


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