Cooperative binding of TEF-1 to repeated GGAATG-related consensus elements with restricted spatial separation and orientation

S. W. Jiang, D. Desai, S. Khan, N. L. Eberhardt

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The human transcriptional enhancer factor (TEF) family includes TEF-1, TEF-3, TEF-4, and TEF-5. The TEFs share a highly conserved 68-amino acid TEA/ATTS DNA-binding domain, which binds to SV40 GT-IIC (GGAATG), SphI (AGTATG), SphII (AGCATG), and muscle-specific M-CAT (GGTATG) enhansons. We determined the optimal DNA-binding consensus sequence for TEF-1. Using a purified GST-TEF-1 fusion protein and a random pool of synthetic oligonucleotides, 31 independent clones were obtained after six rounds of binding site selection. DNA sequences analysis revealed that 16 clones contained direct repeats with a 3-bp spacer (DR3), and 15 clones contained a single binding site. The predominate consensus half-site was GGAATG (67%), and the other elements were of the form GAGAT/CATG. The TEF-1 bound to the DR3 as a dimer in a cooperative manner. Cooperative binding was dependent on the spacing and orientation of the half-sites and was inhibited by deoxycholate treatment, providing evidence that protein-protein interactions were involved. The data suggest that TEF dimerization is important for its ability to modulate gene transcription.

Original languageEnglish (US)
Pages (from-to)507-514
Number of pages8
JournalDNA and Cell Biology
Volume19
Issue number8
DOIs
StatePublished - 2000

Fingerprint

Clone Cells
Binding Sites
Proteins
Deoxycholic Acid
Nucleic Acid Repetitive Sequences
DNA
Consensus Sequence
Dimerization
DNA Sequence Analysis
Oligonucleotides
Amino Acids
Muscles
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Cooperative binding of TEF-1 to repeated GGAATG-related consensus elements with restricted spatial separation and orientation. / Jiang, S. W.; Desai, D.; Khan, S.; Eberhardt, N. L.

In: DNA and Cell Biology, Vol. 19, No. 8, 2000, p. 507-514.

Research output: Contribution to journalArticle

Jiang, S. W. ; Desai, D. ; Khan, S. ; Eberhardt, N. L. / Cooperative binding of TEF-1 to repeated GGAATG-related consensus elements with restricted spatial separation and orientation. In: DNA and Cell Biology. 2000 ; Vol. 19, No. 8. pp. 507-514.
@article{eaca66a333c94737a37a09e6f3f2ad4e,
title = "Cooperative binding of TEF-1 to repeated GGAATG-related consensus elements with restricted spatial separation and orientation",
abstract = "The human transcriptional enhancer factor (TEF) family includes TEF-1, TEF-3, TEF-4, and TEF-5. The TEFs share a highly conserved 68-amino acid TEA/ATTS DNA-binding domain, which binds to SV40 GT-IIC (GGAATG), SphI (AGTATG), SphII (AGCATG), and muscle-specific M-CAT (GGTATG) enhansons. We determined the optimal DNA-binding consensus sequence for TEF-1. Using a purified GST-TEF-1 fusion protein and a random pool of synthetic oligonucleotides, 31 independent clones were obtained after six rounds of binding site selection. DNA sequences analysis revealed that 16 clones contained direct repeats with a 3-bp spacer (DR3), and 15 clones contained a single binding site. The predominate consensus half-site was GGAATG (67{\%}), and the other elements were of the form GAGAT/CATG. The TEF-1 bound to the DR3 as a dimer in a cooperative manner. Cooperative binding was dependent on the spacing and orientation of the half-sites and was inhibited by deoxycholate treatment, providing evidence that protein-protein interactions were involved. The data suggest that TEF dimerization is important for its ability to modulate gene transcription.",
author = "Jiang, {S. W.} and D. Desai and S. Khan and Eberhardt, {N. L.}",
year = "2000",
doi = "10.1089/10445490050128430",
language = "English (US)",
volume = "19",
pages = "507--514",
journal = "DNA and Cell Biology",
issn = "1044-5498",
publisher = "Mary Ann Liebert Inc.",
number = "8",

}

TY - JOUR

T1 - Cooperative binding of TEF-1 to repeated GGAATG-related consensus elements with restricted spatial separation and orientation

AU - Jiang, S. W.

AU - Desai, D.

AU - Khan, S.

AU - Eberhardt, N. L.

PY - 2000

Y1 - 2000

N2 - The human transcriptional enhancer factor (TEF) family includes TEF-1, TEF-3, TEF-4, and TEF-5. The TEFs share a highly conserved 68-amino acid TEA/ATTS DNA-binding domain, which binds to SV40 GT-IIC (GGAATG), SphI (AGTATG), SphII (AGCATG), and muscle-specific M-CAT (GGTATG) enhansons. We determined the optimal DNA-binding consensus sequence for TEF-1. Using a purified GST-TEF-1 fusion protein and a random pool of synthetic oligonucleotides, 31 independent clones were obtained after six rounds of binding site selection. DNA sequences analysis revealed that 16 clones contained direct repeats with a 3-bp spacer (DR3), and 15 clones contained a single binding site. The predominate consensus half-site was GGAATG (67%), and the other elements were of the form GAGAT/CATG. The TEF-1 bound to the DR3 as a dimer in a cooperative manner. Cooperative binding was dependent on the spacing and orientation of the half-sites and was inhibited by deoxycholate treatment, providing evidence that protein-protein interactions were involved. The data suggest that TEF dimerization is important for its ability to modulate gene transcription.

AB - The human transcriptional enhancer factor (TEF) family includes TEF-1, TEF-3, TEF-4, and TEF-5. The TEFs share a highly conserved 68-amino acid TEA/ATTS DNA-binding domain, which binds to SV40 GT-IIC (GGAATG), SphI (AGTATG), SphII (AGCATG), and muscle-specific M-CAT (GGTATG) enhansons. We determined the optimal DNA-binding consensus sequence for TEF-1. Using a purified GST-TEF-1 fusion protein and a random pool of synthetic oligonucleotides, 31 independent clones were obtained after six rounds of binding site selection. DNA sequences analysis revealed that 16 clones contained direct repeats with a 3-bp spacer (DR3), and 15 clones contained a single binding site. The predominate consensus half-site was GGAATG (67%), and the other elements were of the form GAGAT/CATG. The TEF-1 bound to the DR3 as a dimer in a cooperative manner. Cooperative binding was dependent on the spacing and orientation of the half-sites and was inhibited by deoxycholate treatment, providing evidence that protein-protein interactions were involved. The data suggest that TEF dimerization is important for its ability to modulate gene transcription.

UR - http://www.scopus.com/inward/record.url?scp=0034470520&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034470520&partnerID=8YFLogxK

U2 - 10.1089/10445490050128430

DO - 10.1089/10445490050128430

M3 - Article

C2 - 10975468

AN - SCOPUS:0034470520

VL - 19

SP - 507

EP - 514

JO - DNA and Cell Biology

JF - DNA and Cell Biology

SN - 1044-5498

IS - 8

ER -