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dc.contributor.authorBoyd, Aoife
dc.contributor.authorAbram, Florence
dc.contributor.authorKaratzas, Kimon-andreas
dc.contributor.authorMatlawska-Wasowska, Ksenia
dc.contributor.authorConnolly, David
dc.contributor.authorO'Byrne, Conor P.
dc.contributor.authorStarr, Emily
dc.date.accessioned2013-11-29T12:29:05Z
dc.date.available2013-11-29T12:29:05Z
dc.date.issued2008-09
dc.identifier.citationAbram, F,Starr, E,Karatzas, KAG,Matlawska-Wasowska, K,Boyd, A,Wiedmann, M,Boor, KJ,Connally, D,O'Byrne, CP (2008) 'Identification of Components of the Sigma B Regulon in Listeria monocytogenes That Contribute to Acid and Salt Tolerance'. Applied And Environmental Microbiology, 74 :6848-6858.en_US
dc.identifier.urihttp://hdl.handle.net/10379/3859
dc.description.abstractSigma B is an alternative sigma factor that controls the transcriptional response to stress in Listeria monocytogenes and is also known to play a role in the virulence of this human pathogen. In the present study we investigated the impact of a sigB deletion on the proteome of L. monocytogenes grown in a chemically defined medium both in the presence and in the absence of osmotic stress (0.5 M NaCl). Two new phenotypes associated with the sigB deletion were identified using this medium. (i) Unexpectedly, the strain with the Delta sigB deletion was found to grow faster than the parent strain in the growth medium, but only when 0.5 M NaCl was present. This phenomenon was independent of the carbon source provided in the medium. (ii) The Delta sigB mutant was found to have unusual Gram staining properties compared to the parent, suggesting that sigma(B) contributes to the maintenance of an intact cell wall. A proteomic analysis was performed by two-dimensional gel electrophoresis, using cells growing in the exponential and stationary phases. Overall, 11 proteins were found to be differentially expressed in the wild type and the Delta sigB mutant; 10 of these proteins were expressed at lower levels in the mutant, and 1 was overexpressed in the mutant. All 11 proteins were identified by tandem mass spectrometry, and putative functions were assigned based on homology to proteins from other bacteria. Five proteins had putative functions related to carbon utilization (Lmo0539, Lmo0783, Lmo0913, Lmo1830, and Lmo2696), while three proteins were similar to proteins whose functions are unknown but that are known to be stress inducible (Lmo0796, Lmo2391, and Lmo2748). To gain further insight into the role of sigma(B) in L. monocytogenes, we deleted the genes encoding four of the proteins, lmo0796, lmo0913, lmo2391, and lmo2748. Phenotypic characterization of the mutants revealed that Lmo2748 plays a role in osmotolerance, while Lmo0796, Lmo0913, and Lmo2391 were all implicated in acid stress tolerance to various degrees. Invasion assays performed with Caco-2 cells indicated that none of the four genes was required for mammalian cell invasion. Microscopic analysis suggested that loss of Lmo2748 might contribute to the cell wall defect observed in the Delta sigB mutant. Overall, this study highlighted two new phenotypes associated with the loss of sigma(B). It also demonstrated clear roles for sigma(B) in both osmotic and low-pH stress tolerance and identified specific components of the sigma(B) regulon that contribute to the responses observed.en_US
dc.formatapplication/pdfen_US
dc.language.isoenen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.relation.ispartofApplied And Environmental Microbiologyen
dc.subjectGENERAL STRESS-RESPONSEen_US
dc.subjectBACILLUS-SUBTILISen_US
dc.subjectSTAPHYLOCOCCUS-AUREUSen_US
dc.subjectESCHERICHIA-COLIen_US
dc.subjectGLUTAMATE-DECARBOXYLASEen_US
dc.subjectSTARVATION SURVIVALen_US
dc.subjectFACTOR COMPETITIONen_US
dc.subjectRPOS MUTATIONSen_US
dc.subjectRNA-POLYMERASEen_US
dc.subjectGROWTHen_US
dc.titleIdentification of Components of the Sigma B Regulon in Listeria monocytogenes That Contribute to Acid and Salt Toleranceen_US
dc.typeArticleen_US
dc.date.updated2013-09-20T14:15:39Z
dc.identifier.doi10.1128/AEM.00442-08
dc.local.publishedsourcehttp://dx.doi.org/10.1128/AEM.00442-08en_US
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funderNational Institutes of Health award RO1-AI052151-01A1 (to K.J.B.)
dc.contributor.funderScience Foundation Ireland Research Frontiers Programme
dc.contributor.funderFramework 6 Marie Curie Transfer of Knowledge grant
dc.internal.rssid1334849
dc.local.contactAoife Boyd, Dept. Of Microbiology, Arts/Science Building, Nui Galway. 2404 Email: aoife.boyd@nuigalway.ie
dc.local.copyrightcheckedNo
dc.local.versionPUBLISHED
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