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PMID: 8855228 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

High-resolution mapping of nucleoprotein complexes by site-specific protein-DNA photocrosslinking: organization of the human TBP-TFIIA-TFIIB-DNA quaternary complex.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 20 ·1996-10-01 ·Pages 10620-5

Lagrange T, Kim TK, Orphanides G, Ebright YW, Ebright RH, Reinberg D

Abstract

We have used a novel site-specific protein-DNA photocrosslinking procedure to define the positions of polypeptide chains relative to promoter DNA in binary, ternary, and quaternary complexes containing human TATA-binding protein, human or yeast transcription factor IIA (TFIIA), human transcription factor IIB (TFIIB), and promoter DNA. The results indicate that TFIIA and TFIIB make more extensive interactions with promoter DNA than previously anticipated. TATA-binding protein, TFIIA, and TFIIB surround promoter DNA for two turns of DNA helix and thus may form a "cylindrical clamp" effectively topologically linked to promoter DNA. Our results have implications for the energetics, DNA-sequence-specificity, and pathway of assembly of eukaryotic transcription complexes.

MeSH Terms
Adenoviruses, Human Base Sequence Cross-Linking Reagents DNA, Viral/ultrastructure DNA-Binding Proteins/chemistry,ultrastructure Deoxyribonucleoproteins/ultrastructure Fungal Proteins/chemistry,ultrastructure Humans Macromolecular Substances Photochemistry Promoter Regions, Genetic Saccharomyces cerevisiae TATA-Box Binding Protein Transcription Factor TFIIA Transcription Factor TFIIB Transcription Factors/chemistry,ultrastructure
Chemicals
Cross-Linking Reagents DNA, Viral DNA-Binding Proteins Deoxyribonucleoproteins Fungal Proteins Macromolecular Substances TATA-Box Binding Protein Transcription Factor TFIIA Transcription Factor TFIIB Transcription Factors
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lagrange T
Howard Hughes Medical Institute, Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854, USA.
Kim T K
Orphanides G
Ebright Y W
Ebright R H
Reinberg D
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.20.10620
Published
1996-10-01
Pages
10620-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC38203
Subset
IM
Grants
NIGMS NIH HHS · GM37120 · United States
NIGMS NIH HHS · GM47484 · United States
NIGMS NIH HHS · GM48518 · United States
External Links
PubMed source DOI: 10.1073/pnas.93.20.10620 PMC Full Text View journal details
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