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Vibrio parahaemolyticus  (Vp) is a marine halophilic bacterium that is commonly associated with oysters and shrimp. Human consumption of contaminated shellfish can result in Vp mediated gastroenteritis and severe diarrheal disease. Vp encodes two type 3 secretion systems (T3SS-1 and T3SS2) that have been functionally implicated in cytotoxicity and enterotoxicity respectively. In this study, we profiled protein secretion and temporal promoter activities associated with  exsA  and  exsB  gene expression.  exsA  is an AraC-like transcriptional activator that is critical for activating multiple operons that encode T3SS-1 genes, whereas  exsB  is thought to encode an outer membrane pilotin component for T3SS-1. The  exsBA  genetic locus has two predicted promoter elements. The predicted  exsB  and  exsA  promoters were individually cloned upstream of  luxCDABE  genes in reporter plasmid constructs allowing for  in situ , real-time quantitative light emission measurements under many growth conditions. Low calcium growth conditions supported maximal  exsB  and  exsA  promoter activation.  exsB  promoter activity exhibited high basal activity and resulted in an  exsBA  co-transcript. Furthermore, a separate proximal  exsA  promoter showed initial low basal activity yet eventually exceeded that of  exsB  and reached maximal levels after 2.5 h corresponding to an entry into early log phase.  exsA  promoter activity was significantly higher at 30°C than 37°C, which also coincided with increased secretion levels of specific T3SS-1 effector proteins. Lastly, bioinformatic analyses identified a putative expanded ExsA binding motif for multiple transcriptional operons. These findings suggest a two wave model of Vp T3SS-I induction that integrates two distinct promoter elements and environmental signals into a complex ExsA activation framework.

Transcriptional profiling of Vibrio parahaemolyticus exsA reveals a complex activation network for type III secretion