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PMID: 8306968 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The chromatin-associated protein H-NS alters DNA topology in vitro.

The EMBO journal ·Vol. 13 ·No. 1 ·1994-01-01 ·Pages 258-68

Tupper AE, Owen-Hughes TA, Ussery DW, Santos DS, Ferguson DJ, Sidebotham JM, Hinton JC, Higgins CF

Abstract

H-NS is one of the two most abundant proteins in the bacterial nucleoid and influences the expression of a number of genes. We have studied the interaction of H-NS with DNA; purified H-NS was demonstrated to constrain negative DNA supercoils in vitro. This provides support for the hypothesis that H-NS influences transcription via changes in DNA topology, and is evidence of a structural role for H-NS in bacterial chromatin. The effects of H-NS on topology were only observed at sub-saturating concentrations of the protein. In addition, a preferred binding site on DNA was identified by DNase I footprinting at sub-saturating H-NS concentrations. This site corresponded to a curved sequence element which we previously showed, by in vivo studies, to be a site at which H-NS influences transcription of the proU operon. When present in saturating concentrations, H-NS did not constrain supercoils and bound to DNA in a sequence-independent fashion, covering all DNA molecules from end to end, suggesting that H-NS may form distinct complexes with DNA at different H-NS:DNA ratios. The data presented here provide direct support for the hypothesis that H-NS acts at specific sites to influence DNA topology and, hence, transcription.

MeSH Terms
Bacterial Proteins Base Sequence Chromatin/metabolism DNA, Bacterial/chemistry,metabolism,ultrastructure DNA-Binding Proteins/metabolism,ultrastructure Deoxyribonuclease I Molecular Sequence Data Nuclear Proteins/metabolism,ultrastructure Nucleic Acid Conformation Osmolar Concentration Promoter Regions, Genetic Protein Binding Salmonella typhimurium Transcription, Genetic
Chemicals
Bacterial Proteins Chromatin DNA, Bacterial DNA-Binding Proteins H-NS protein, bacteria Nuclear Proteins Deoxyribonuclease I
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Tupper A E
Imperial Cancer Research Fund Laboratories, University of Oxford, John Radcliffe Hospital, UK.
Owen-Hughes T A
Ussery D W
Santos D S
Ferguson D J
Sidebotham J M
Hinton J C
Higgins C F
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-01-01
Pages
258-68
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394800
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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