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PMID: 9843508 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Thermoregulation of Shigella and Escherichia coli EIEC pathogenicity. A temperature-dependent structural transition of DNA modulates accessibility of virF promoter to transcriptional repressor H-NS.

The EMBO journal ·Vol. 17 ·No. 23 ·1998-12-01 ·Pages 7033-43

Falconi M, Colonna B, Prosseda G, Micheli G, Gualerzi CO

Abstract

The expression of plasmid-borne virF of Shigella encoding a transcriptional regulator of the AraC family, is required to initiate a cascade of events resulting in activation of several operons encoding invasion functions. H-NS, one of the main nucleoid-associated proteins, controls the temperature-dependent expression of the virulence genes by repressing the in vivo transcription of virF only below a critical temperature (approximately 32 degrees C). This temperature-dependent transcriptional regulation has been reproduced in vitro and the targets of H-NS on the virF promoter were identified as two sites centred around -250 and -1 separated by an intrinsic DNA curvature. H-NS bound cooperatively to these two sites below 32 degrees C, but not at 37 degrees C. DNA supercoiling within the virF promoter region did not influence H-NS binding but was necessary for the H-NS-mediated transcriptional repression. Electrophoretic analysis between 4 and 60 degrees C showed that the virF promoter fragment, comprising the two H-NS sites, undergoes a specific and temperature-dependent conformational transition at approximately 32 degrees C. Our results suggest that this modification of the DNA target may modulate a cooperative interaction between H-NS molecules bound at two distant sites in the virF promoter region and thus represents the physical basis for the H-NS-dependent thermoregulation of virulence gene expression.

MeSH Terms
Bacterial Proteins/genetics Base Sequence DNA, Bacterial DNA-Binding Proteins/metabolism Escherichia coli/genetics,pathogenicity Gene Expression Regulation, Bacterial Molecular Sequence Data Promoter Regions, Genetic Repressor Proteins/metabolism Shigella flexneri/genetics,pathogenicity Temperature Transcription, Genetic Virulence Factors
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins H-NS protein, bacteria Repressor Proteins Virulence Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Falconi M
Laboratorio di Genetica, Dipartimento di Biologia MCA, Università di Camerino, 62032 Camerino (MC), Italy.
Colonna B
Prosseda G
Micheli G
Gualerzi C O
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.23.7033
Published
1998-12-01
Pages
7033-43
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171051
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.23.7033 PMC Full Text View journal details
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