Regulation of type III secretion system 1 gene expression in Vibrio parahaemolyticus is dependent on interactions between ExsA, ExsC, and ExsD

Xiaohui Zhou; Michael E Konkel; Douglas R Call

doi:10.4161/viru.1.4.12318

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Regulation of type III secretion system 1 gene expression in Vibrio parahaemolyticus is dependent on interactions between ExsA, ExsC, and ExsD

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Regulation of type III secretion system 1 gene expression in Vibrio parahaemolyticus is dependent on interactions between ExsA, ExsC, and ExsD

Xiaohui Zhou, Michael E Konkel and Douglas R Call

Virulence, Vol.1(4), pp.260-272

07/2010

DOI: https://doi.org/10.4161/viru.1.4.12318

Handle:

https://hdl.handle.net/2376/101741

PMCID: PMC3073295

PMID: 21178451

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Abstract

Repressor Proteins - chemistry

Humans

Bacterial Proteins - chemistry

Bacterial Proteins - genetics

Repressor Proteins - genetics

Vibrio parahaemolyticus - metabolism

Trans-Activators - chemistry

Recombinant Fusion Proteins - metabolism

Vibrio parahaemolyticus - genetics

Culture Media

Trans-Activators - genetics

Protein Binding

Recombinant Fusion Proteins - genetics

Bacterial Proteins - metabolism

Trans-Activators - metabolism

Transcription, Genetic

Vibrio parahaemolyticus - growth & development

HeLa Cells

Gene Expression Regulation, Bacterial

Repressor Proteins - metabolism

Vibrio parahaemolyticus ExsA is the transcriptional regulator for type III secretion system 1 (T3SS1) while ExsD blocks T3SS1 expression. Herein we show that deletion of exsC from V. parahaemolyticus blocked synthesis of T3SS1-dependent proteins under inducing conditions (contact with HeLa cells), while in trans complementation of the ΔexsC strain with wild-type exsC restored protein synthesis. Under non-inducing conditions (Luria broth plus salt), in trans expression of exsC in a wild-type strain resulted in synthesis and secretion of T3SS1-dependent proteins. Deletion of exsC does not affect the synthesis of ExsA while expression of T3SS1 genes is independent of ExsC in the absence of ExsD. Co-expression of recombinant proteins with different antigenic tags demonstrated that ExsC binds ExsD and that the N-terminal amino acids of ExsC (positions 7 to 12) are required for binding. Co-expression and purification of antigentically tagged ExsA and ExsD demonstrated that ExsD directly binds ExsA and presumably prevents ExsA from binding promoter regions of T3SS1 genes. Collectively these data demonstrate that ExsD binds ExsA to block expression of T3SS1 genes, while ExsC binds ExsD to permit expression of T3SS1 genes. ExsA, ExsC, and ExsD from V. parahaemolyticus appear to be functional orthologues of their Pseudomonas aeruginosa counterparts.

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Title: Regulation of type III secretion system 1 gene expression in Vibrio parahaemolyticus is dependent on interactions between ExsA, ExsC, and ExsD
Creators: Xiaohui Zhou - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA
Michael E Konkel
Douglas R Call
Publication Details: Virulence, Vol.1(4), pp.260-272
Academic Unit: Paul G. Allen School for Global Animal Health; Molecular Biosciences, School of
Publisher: United States
Grant note: N01AI30055 / NIAID NIH HHS
Identifiers: 99900546557701842
Language: English
Resource Type: Journal article