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Interaction of the Salmonella typhimurium transcription and virulence factor SlyA with target DNA and identification of members of the SlyA regulon

Interaction of the Salmonella typhimurium transcription and virulence factor SlyA with target DNA and identification of members of the SlyA regulon
Interaction of the Salmonella typhimurium transcription and virulence factor SlyA with target DNA and identification of members of the SlyA regulon

The SlyA protein from Salmonella typhimurium is a transcription factor that contributes to virulence. It is shown that a slyA mutant is attenuated in the presence of murine macrophages compared with the parent strain. Moreover, after growth in minimal medium, survival of the slyA mutant was reduced. Altered levels of flagellin (fliC), PagC, IroN, and outer membrane proteins suggest that the slyA mutation affects the surface properties of Salmonella. The isolated SlyA protein is a cofactor-free homodimer that recognizes five sites within the promoter region of the slyA gene. One of these sites contained a near perfect inverted repeat TTAGCAAGCTAA. The other four sites contained related sequences. Occupation of the SlyA sites in the slyA promoter prevented open-complex formation, consistent with the pattern of slyA::lacZ expression parental and slyA mutant strains. By combining the footprinting data with potential SlyA binding sites recovered from a pool of random DNA sequences, a consensus was defined and used to probe the NIH Salmonella unfinished genomes data base. These searches revealed the presence of consensus SlyA sites upstream of omp, ispA, xseB, slyA, and a gene encoding a protein with homology to a hemagglutinin. Accordingly, transcription of an omp::lacZ fusion was reduced in a slyA mutant. Given the difficulties in obtaining a comprehensive picture of intracellular gene expression, the definition of the DNA sequence recognized by a transcription factor (SlyA) that is essential for survival in the macrophage environment should allow a complete regulon of genes with altered expression upon exposure to macrophages to be determined once the S. typhimurium genome annotation is complete.

Animals, Bacterial Proteins, Bacterial Toxins, Base Sequence, Binding Sites, DNA, Bacterial, Electrophoresis, Polyacrylamide Gel, Flagellin, Hemolysin Proteins, Macrophages, Membrane Proteins, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Promoter Regions, Genetic, Regulon, Salmonella typhimurium, Transcription Factors, Transcription, Genetic, Journal Article, Research Support, Non-U.S. Gov't
0021-9258
17630-17637
Stapleton, Melanie R
f0e937d3-51dc-4b00-b909-4c8aa41afd8c
Norte, Valia A
6a5be46e-292c-485f-973b-05d4edfaecaa
Read, Robert C
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Green, Jeffrey
bd38753d-70c3-4327-9893-090ffb2b15ea
Stapleton, Melanie R
f0e937d3-51dc-4b00-b909-4c8aa41afd8c
Norte, Valia A
6a5be46e-292c-485f-973b-05d4edfaecaa
Read, Robert C
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Green, Jeffrey
bd38753d-70c3-4327-9893-090ffb2b15ea

Stapleton, Melanie R, Norte, Valia A, Read, Robert C and Green, Jeffrey (2002) Interaction of the Salmonella typhimurium transcription and virulence factor SlyA with target DNA and identification of members of the SlyA regulon. The Journal of Biological Chemistry, 277 (20), 17630-17637. (doi:10.1074/jbc.M110178200).

Record type: Article

Abstract

The SlyA protein from Salmonella typhimurium is a transcription factor that contributes to virulence. It is shown that a slyA mutant is attenuated in the presence of murine macrophages compared with the parent strain. Moreover, after growth in minimal medium, survival of the slyA mutant was reduced. Altered levels of flagellin (fliC), PagC, IroN, and outer membrane proteins suggest that the slyA mutation affects the surface properties of Salmonella. The isolated SlyA protein is a cofactor-free homodimer that recognizes five sites within the promoter region of the slyA gene. One of these sites contained a near perfect inverted repeat TTAGCAAGCTAA. The other four sites contained related sequences. Occupation of the SlyA sites in the slyA promoter prevented open-complex formation, consistent with the pattern of slyA::lacZ expression parental and slyA mutant strains. By combining the footprinting data with potential SlyA binding sites recovered from a pool of random DNA sequences, a consensus was defined and used to probe the NIH Salmonella unfinished genomes data base. These searches revealed the presence of consensus SlyA sites upstream of omp, ispA, xseB, slyA, and a gene encoding a protein with homology to a hemagglutinin. Accordingly, transcription of an omp::lacZ fusion was reduced in a slyA mutant. Given the difficulties in obtaining a comprehensive picture of intracellular gene expression, the definition of the DNA sequence recognized by a transcription factor (SlyA) that is essential for survival in the macrophage environment should allow a complete regulon of genes with altered expression upon exposure to macrophages to be determined once the S. typhimurium genome annotation is complete.

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More information

Published date: 17 May 2002
Keywords: Animals, Bacterial Proteins, Bacterial Toxins, Base Sequence, Binding Sites, DNA, Bacterial, Electrophoresis, Polyacrylamide Gel, Flagellin, Hemolysin Proteins, Macrophages, Membrane Proteins, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Promoter Regions, Genetic, Regulon, Salmonella typhimurium, Transcription Factors, Transcription, Genetic, Journal Article, Research Support, Non-U.S. Gov't

Identifiers

Local EPrints ID: 416548
URI: http://eprints.soton.ac.uk/id/eprint/416548
ISSN: 0021-9258
PURE UUID: 792731f5-7c23-4a46-b6d9-a2be06e0104c
ORCID for Robert C Read: ORCID iD orcid.org/0000-0002-4297-6728

Catalogue record

Date deposited: 21 Dec 2017 17:30
Last modified: 10 Nov 2021 03:30

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Contributors

Author: Melanie R Stapleton
Author: Valia A Norte
Author: Robert C Read ORCID iD
Author: Jeffrey Green

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