Simultaneous binding and bending of promoter DNA by the TATA binding protein

Real time kinetic measurements

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

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

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
JournalBiochemistry
Volume35
Issue number23
DOIs
StatePublished - Jun 11 1996

Fingerprint

TATA-Box Binding Protein
DNA-Binding Proteins
Fluorescein
Kinetics
DNA
TATA Box
Rhodamines
X ray crystallography
Saccharomyces cerevisiae Proteins
Oligomers
Fluorometry
Yeast
Probability distributions
Fluorescence Resonance Energy Transfer
Rate constants
X Ray Crystallography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Simultaneous binding and bending of promoter DNA by the TATA binding protein : Real time kinetic measurements. / Parkhurst, Kay M.; Brenowitz, Michael D.; Parkhurst, Lawrence J.

In: Biochemistry, Vol. 35, No. 23, 11.06.1996, p. 7459-7465.

Research output: Contribution to journalArticle

@article{1c628566d2f64268b836ba99054338ea,
title = "Simultaneous binding and bending of promoter DNA by the TATA binding protein: Real time kinetic measurements",
abstract = "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 {\AA} 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.",
author = "Parkhurst, {Kay M.} and Brenowitz, {Michael D.} and Parkhurst, {Lawrence J.}",
year = "1996",
month = "6",
day = "11",
doi = "10.1021/bi9530301",
language = "English (US)",
volume = "35",
pages = "7459--7465",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "23",

}

TY - JOUR

T1 - Simultaneous binding and bending of promoter DNA by the TATA binding protein

T2 - Real time kinetic measurements

AU - Parkhurst, Kay M.

AU - Brenowitz, Michael D.

AU - Parkhurst, Lawrence J.

PY - 1996/6/11

Y1 - 1996/6/11

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0029943025&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029943025&partnerID=8YFLogxK

U2 - 10.1021/bi9530301

DO - 10.1021/bi9530301

M3 - Article

VL - 35

SP - 7459

EP - 7465

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 23

ER -