Dimerization of the TATA binding protein

Robert A. Coleman, A. K P Taggart, L. R. Benjamin, B. F. Pugh

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

The TATA binding protein (TBP) is a central component of all eukaryotic transcription machineries. The recruitment of TBP to the promoter is slow and possibly rate limiting in transcription complex assembly. In an effort to understand the nature of this potential rate-limiting step, we have investigated the physical state of TBP prior to DNA binding. By chemical cross-linking, gel filtration chromatography, and protein affinity chromatography, we find that the conserved carboxyl-terminal DNA binding domain of human TBP dimerizes when not bound to DNA. The data completely support the proposed dimeric structure of plant TBP, previously determined by x-ray crystallography. TBP dimers are quite stable, having an approximate equilibrium dissociation constant (K(D)) in the low nanomolar range. The dimerization interface appears to be dominated by hydrophobic forces, as predicted by the crystal structure. TBP dimers do not bind DNA, but they must dissociate into monomers before stably binding to the TATA box. Dissociation of TBP dimers appears to be relatively slow, and as such has the potential to dictate the kinetics of DNA binding.

Original languageEnglish (US)
Pages (from-to)13842-13849
Number of pages8
JournalJournal of Biological Chemistry
Volume270
Issue number23
DOIs
StatePublished - 1995
Externally publishedYes

Fingerprint

TATA-Box Binding Protein
Dimerization
Dimers
DNA
Transcription
Affinity chromatography
Plant Proteins
TATA Box
Crystallography
Chromatography
Affinity Chromatography
Gel Chromatography
Monomers
Crystal structure
Gels
X-Rays
X rays
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Coleman, R. A., Taggart, A. K. P., Benjamin, L. R., & Pugh, B. F. (1995). Dimerization of the TATA binding protein. Journal of Biological Chemistry, 270(23), 13842-13849. https://doi.org/10.1074/jbc.270.23.13842

Dimerization of the TATA binding protein. / Coleman, Robert A.; Taggart, A. K P; Benjamin, L. R.; Pugh, B. F.

In: Journal of Biological Chemistry, Vol. 270, No. 23, 1995, p. 13842-13849.

Research output: Contribution to journalArticle

Coleman, RA, Taggart, AKP, Benjamin, LR & Pugh, BF 1995, 'Dimerization of the TATA binding protein', Journal of Biological Chemistry, vol. 270, no. 23, pp. 13842-13849. https://doi.org/10.1074/jbc.270.23.13842
Coleman, Robert A. ; Taggart, A. K P ; Benjamin, L. R. ; Pugh, B. F. / Dimerization of the TATA binding protein. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 23. pp. 13842-13849.
@article{f5270a5f1eeb444cb05c6b8978eb387d,
title = "Dimerization of the TATA binding protein",
abstract = "The TATA binding protein (TBP) is a central component of all eukaryotic transcription machineries. The recruitment of TBP to the promoter is slow and possibly rate limiting in transcription complex assembly. In an effort to understand the nature of this potential rate-limiting step, we have investigated the physical state of TBP prior to DNA binding. By chemical cross-linking, gel filtration chromatography, and protein affinity chromatography, we find that the conserved carboxyl-terminal DNA binding domain of human TBP dimerizes when not bound to DNA. The data completely support the proposed dimeric structure of plant TBP, previously determined by x-ray crystallography. TBP dimers are quite stable, having an approximate equilibrium dissociation constant (K(D)) in the low nanomolar range. The dimerization interface appears to be dominated by hydrophobic forces, as predicted by the crystal structure. TBP dimers do not bind DNA, but they must dissociate into monomers before stably binding to the TATA box. Dissociation of TBP dimers appears to be relatively slow, and as such has the potential to dictate the kinetics of DNA binding.",
author = "Coleman, {Robert A.} and Taggart, {A. K P} and Benjamin, {L. R.} and Pugh, {B. F.}",
year = "1995",
doi = "10.1074/jbc.270.23.13842",
language = "English (US)",
volume = "270",
pages = "13842--13849",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "23",

}

TY - JOUR

T1 - Dimerization of the TATA binding protein

AU - Coleman, Robert A.

AU - Taggart, A. K P

AU - Benjamin, L. R.

AU - Pugh, B. F.

PY - 1995

Y1 - 1995

N2 - The TATA binding protein (TBP) is a central component of all eukaryotic transcription machineries. The recruitment of TBP to the promoter is slow and possibly rate limiting in transcription complex assembly. In an effort to understand the nature of this potential rate-limiting step, we have investigated the physical state of TBP prior to DNA binding. By chemical cross-linking, gel filtration chromatography, and protein affinity chromatography, we find that the conserved carboxyl-terminal DNA binding domain of human TBP dimerizes when not bound to DNA. The data completely support the proposed dimeric structure of plant TBP, previously determined by x-ray crystallography. TBP dimers are quite stable, having an approximate equilibrium dissociation constant (K(D)) in the low nanomolar range. The dimerization interface appears to be dominated by hydrophobic forces, as predicted by the crystal structure. TBP dimers do not bind DNA, but they must dissociate into monomers before stably binding to the TATA box. Dissociation of TBP dimers appears to be relatively slow, and as such has the potential to dictate the kinetics of DNA binding.

AB - The TATA binding protein (TBP) is a central component of all eukaryotic transcription machineries. The recruitment of TBP to the promoter is slow and possibly rate limiting in transcription complex assembly. In an effort to understand the nature of this potential rate-limiting step, we have investigated the physical state of TBP prior to DNA binding. By chemical cross-linking, gel filtration chromatography, and protein affinity chromatography, we find that the conserved carboxyl-terminal DNA binding domain of human TBP dimerizes when not bound to DNA. The data completely support the proposed dimeric structure of plant TBP, previously determined by x-ray crystallography. TBP dimers are quite stable, having an approximate equilibrium dissociation constant (K(D)) in the low nanomolar range. The dimerization interface appears to be dominated by hydrophobic forces, as predicted by the crystal structure. TBP dimers do not bind DNA, but they must dissociate into monomers before stably binding to the TATA box. Dissociation of TBP dimers appears to be relatively slow, and as such has the potential to dictate the kinetics of DNA binding.

UR - http://www.scopus.com/inward/record.url?scp=0029013642&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029013642&partnerID=8YFLogxK

U2 - 10.1074/jbc.270.23.13842

DO - 10.1074/jbc.270.23.13842

M3 - Article

VL - 270

SP - 13842

EP - 13849

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 23

ER -