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Type III secretion system 1 genes in Vibrio parahaemolyticus are positively regulated by ExsA and negatively regulated by ExsD.

ABSTRACT: Vibrio parahaemolyticus harbours two distinct type III secretion systems (T3SS1 and T3SS2). A subset of 10 T3SS1 genes are transcribed when V. parahaemolyticus is grown in tissue culture medium [Dulbecco's modified Eagle's medium (DMEM)], while transcription of these genes (except exsD) is minimal upon growth in Luria-Bertani-Salt (LB-S). Transcription of T3SS1 genes and cytotoxicity towards HeLa cells was prevented by deletion of exsA while complementation with exsA restored these traits. Overexpression of ExsA in the wild-type strain, NY-4, activated the transcription of T3SS1 genes when bacteria were grown in LB-S. Thus, ExsA is necessary and sufficient to induce the transcription of T3SS1 genes. Deletion of the exsD permitted the transcription of T3SS1 genes when bacteria were grown in the LB-S medium and complementation with the wild-type exsD gene-blocked transcription of T3SS1 genes. Overexpression of ExsD in NY-4 prevented the transcription of T3SS1 gene when bacteria were grown in DMEM. A gel mobility shift assay demonstrated that purified ExsA protein binds a novel motif in the upstream region of vp1668 and vp1687, indicating that ExsA interacts directly with the promoter sequences of T3SS1 genes. ExsA positively regulates the expression and secretion of Vp1656 while ExsD negatively regulates the expression and secretion of Vp1656.

SUBMITTER: Zhou X

PROVIDER: S-EPMC2610376 | biostudies-literature | 2008 Aug

REPOSITORIES: biostudies-literature

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Type III secretion system 1 genes in Vibrio parahaemolyticus are positively regulated by ExsA and negatively regulated by ExsD.

Zhou Xiaohui X Shah Devendra H DH Konkel Michael E ME Call Douglas R DR

Molecular microbiology 20080628 3

Vibrio parahaemolyticus harbours two distinct type III secretion systems (T3SS1 and T3SS2). A subset of 10 T3SS1 genes are transcribed when V. parahaemolyticus is grown in tissue culture medium [Dulbecco's modified Eagle's medium (DMEM)], while transcription of these genes (except exsD) is minimal upon growth in Luria-Bertani-Salt (LB-S). Transcription of T3SS1 genes and cytotoxicity towards HeLa cells was prevented by deletion of exsA while complementation with exsA restored these traits. Overe ...[more]

PMID: 18554322

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Project description:Vibrio parahaemolyticus is a marine bacterium that is globally recognized as the leading cause of seafood-borne gastroenteritis. V. parahaemolyticus uses various toxins and two type 3 secretion systems (T3SS-1 and T3SS-2) to subvert host cells during infection. We previously determined that V. parahaemolyticus T3SS-1 activity is upregulated by increasing the expression level of the master regulator ExsA under specific growth conditions. In this study, we set out to identify V. parahaemolyticus genes responsible for linking environmental and growth signals to exsA gene expression. Using transposon mutagenesis in combination with a sensitive and quantitative luminescence screen, we identify HlyU and H-NS as two antagonistic regulatory proteins controlling the expression of exsA and, hence, T3SS-1 in V. parahaemolyticus Disruption of hns leads to constitutive unregulated exsA gene expression, consistent with its known role in repressing exsA transcription. In contrast, genetic disruption of hlyU completely abrogated exsA expression and T3SS-1 activity. A V. parahaemolyticushlyU null mutant was significantly deficient for T3SS-1-mediated host cell death during in vitro infection. DNA footprinting studies with purified HlyU revealed a 56-bp protected DNA region within the exsA promoter that contains an inverted repeat sequence. Genetic evidence suggests that HlyU acts as a derepressor, likely by displacing H-NS from the exsA promoter, leading to exsA gene expression and appropriately regulated T3SS-1 activity. Overall, the data implicate HlyU as a critical positive regulator of V. parahaemolyticus T3SS-1-mediated pathogenesis.IMPORTANCE Many Vibrio species are zoonotic pathogens, infecting both animals and humans, resulting in significant morbidity and, in extreme cases, mortality. While many Vibrio species virulence genes are known, their associated regulation is often modestly understood. We set out to identify genetic factors of V. parahaemolyticus that are involved in activating exsA gene expression, a process linked to a type III secretion system involved in host cytotoxicity. We discover that V. parahaemolyticus employs a genetic regulatory switch involving H-NS and HlyU to control exsA promoter activity. While HlyU is a well-known positive regulator of Vibrio species virulence genes, this is the first report linking it to a transcriptional master regulator and type III secretion system paradigm.

Dataset Information

Type III secretion system 1 genes in Vibrio parahaemolyticus are positively regulated by ExsA and negatively regulated by ExsD.

Publications

Type III secretion system 1 genes in Vibrio parahaemolyticus are positively regulated by ExsA and negatively regulated by ExsD.

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