Simultaneous binding and bending of promoter DNA by the TATA binding protein: Real time kinetic measurements

Kay M. Parkhurst, Michael Brenowitz, Lawrence J. Parkhurst

Research output: Contribution to journalArticle

116 Scopus citations


The binding and bending of tetramethylrhodamine-5'- (GGGCTATAAAAGGG)(duplex)-3'-fluorescein by native Saccharomyces cerevisiae TATA binding protein (TBP) have been investigated using fluorescence resonance energy transfer. Probability distributions derived from fluorescein emission lifetime measurements show a decrease in the mean 3'-fluorescein- 5'-rhodamine distance from 56.5 to 46.8 Å upon binding of the oligomer to TBP, consistent with the DNA bend observed by X-ray crystallography. The kinetics, monitored in real time using stopped flow fluorimetry, demonstrate simultaneous binding and bending of a TATA box by TBP with a single second- order rate constant of (2.4 ± 0.3) x 106 M-1 s-1 at 30 °C.

Original languageEnglish (US)
Pages (from-to)7459-7465
Number of pages7
Issue number23
StatePublished - Jun 11 1996


ASJC Scopus subject areas

  • Biochemistry

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