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J. Biol. Chem., Vol. 277, Issue 20, 17630-17637, May 17, 2002
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From 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.
Interaction of the Salmonella typhimurium
Transcription and Virulence Factor SlyA with Target DNA and
Identification of Members of the SlyA Regulon*
,,¶
the Krebs Institute for Biomolecular Research,
Department of Molecular Biology and Biotechnology, University of
Sheffield, Western Bank, Sheffield S10 2TN, United Kingdom and the
§ Division of Genomic Medicine, University of Sheffield
Medical School, Sheffield S10 2RX United Kingdom
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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