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The Degree of Oligomerization of the H-NS Nucleoid Structuring Protein Is Related to Specific Binding to DNA

At several E. coli promoters, initiation of transcription is repressed by a tight nucleoprotein complex formed by the assembly of the H-NS protein. In order to characterize the relationship between the structure of H-NS oligomers in solution and on relevant DNA fragments, we have compared wild-type...

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Published in:The Journal of biological chemistry 2002-11, Vol.277 (44), p.41657-41666
Main Authors: Badaut, Cyril, Williams, Roy, Arluison, Veronique, Bouffartigues, Emeline, Robert, Bruno, Buc, Henri, Rimsky, Sylvie
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cited_by cdi_FETCH-LOGICAL-c536t-3c452c20d977c2967f7b83c53da0ecd508dbdc3bcb4fdeae53979747a04f15203
cites cdi_FETCH-LOGICAL-c536t-3c452c20d977c2967f7b83c53da0ecd508dbdc3bcb4fdeae53979747a04f15203
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container_issue 44
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container_title The Journal of biological chemistry
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creator Badaut, Cyril
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Rimsky, Sylvie
description At several E. coli promoters, initiation of transcription is repressed by a tight nucleoprotein complex formed by the assembly of the H-NS protein. In order to characterize the relationship between the structure of H-NS oligomers in solution and on relevant DNA fragments, we have compared wild-type H-NS and several transdominant H-NS mutants using gel shift assays, DNase I footprinting, analytical ultracentrifugation, and reactivity toward a cross-linking reagent. In solution, oligomerization occurs through two protein interfaces, one necessary to construct a dimeric core (and involving residues 1–64) and the other required for subsequent assembly of these dimers. We show that, as well as region 64–95, residues present in the NH 2 -terminal coiled coil domain also participate in this second interface. Our results support the view that the same interacting interfaces are also involved on the DNA. We propose that the dimeric core recognizes specific motifs, with the second interface being critical for their correct head to tail assembly. The COOH-terminal domain of the protein contains the DNA binding motif essential for the discrimination of this specific functional assembly over competitive nonspecific H-NS polymers.
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subjects Amino Acid Transport Systems
Amino Acid Transport Systems - genetics
Bacterial Proteins
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Biochemistry, Molecular Biology
Cross-Linking Reagents
Cross-Linking Reagents - pharmacology
Deoxyribonuclease I
Deoxyribonuclease I - pharmacology
Dimerization
DNA
DNA - chemistry
DNA - metabolism
DNA-Binding Proteins
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - metabolism
Electrophoretic Mobility Shift Assay
Life Sciences
Promoter Regions (Genetics)
Promoter Regions, Genetic
Temperature
Ultracentrifugation
title The Degree of Oligomerization of the H-NS Nucleoid Structuring Protein Is Related to Specific Binding to DNA
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