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PMID: 9286993 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Mutational characterization of promoter regions recognized by the Salmonella dublin virulence plasmid regulatory protein SpvR.

Journal of bacteriology ·Vol. 179 ·No. 17 ·1997-09-00 ·Pages 5398-406

Grob P, Kahn D, Guiney DG

Abstract

The virulence plasmid-encoded spv regulon is essential for virulence of Salmonella dublin in mice. The spvR gene product belongs to the LysR family of transcriptional regulator proteins. SpvR induces the expression of the spvABCD operon and positively regulates its own expression. DNase I protection analysis with purified SpvR fusion proteins identified SpvR binding sites within the spvA and spvR promoters (P. Grob and D. G. Guiney, J. Bacteriol. 178:1813-1820, 1996). We have used PCR mutagenesis, combined with functional selection for reduced SpvR affinity, to define the DNA elements essential for SpvR binding. For the spvR promoter fragment, a screen for reduced expression was also applied. Sequence analysis of the resulting mutant fragments reveals that the base pair changes are clustered in distinct regions. Determination of the apparent dissociation constants of SpvR for the mutant promoters showed that the spvA LysR-type motif and the upstream palindromic sequences of both promoters play an important role in SpvR recognition.

MeSH Terms
Animals Base Sequence Binding Sites DNA Mutational Analysis Genes, Bacterial/genetics Genes, Regulator/genetics Mice Molecular Sequence Data Mutagenesis Plasmids/genetics Promoter Regions, Genetic/genetics Recombinant Fusion Proteins Salmonella/genetics,pathogenicity Transcription Factors/genetics Virulence/genetics
Chemicals
Recombinant Fusion Proteins Transcription Factors
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Grob P
Department of Medicine, School of Medicine, University of California at San Diego, La Jolla 92093-0640, USA.
Kahn D
Guiney D G
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.17.5398-5406.1997
Published
1997-09-00
Pages
5398-406
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179409
Subset
IM
Grants
NIAID NIH HHS · AI-32178 · United States
NIDDK NIH HHS · DK-35108 · United States
External Links
PubMed source DOI: 10.1128/jb.179.17.5398-5406.1997 PMC Full Text View journal details
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