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Transcriptome Analysis of T3SS1 in Vibrio parahaemolyticus

ABSTRACT: Comparative transcriptional mRNA profiles were generated of bacterial pathogen Vibrio parahaemolyticus under conditions that induce activity of virulence factor type III secretion system 1 (T3SS1). Induction conditions included growth of the bacteria in Dulbecco's Modified Eagle Medium (DMEM), overexpression of transcriptional activator exsA and contact with HeLa cells in Hank's Balanced Salt Solution (HBSS), while non-inducing conditions included growth in Luria-Bertani medium supplemented with 2.5% (w/v) NaCl (LB-S) and overexpression of transcriptional anti-activator exsD. Transcriptome profiles of induction conditions were cross-compared against background non-inducing conditions and time points during HeLa cell infection (2 hr, 3 hr, 4 hr, 6 hr, 8 hr) were compared against pre-infection (0 hr) to identify genes important in T3SS1 activity and pathogenesis.

ORGANISM(S): Vibrio parahaemolyticus

PROVIDER: GSE51423 | GEO | 2014/01/21

SECONDARY ACCESSION(S): PRJNA223069

REPOSITORIES: GEO

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Transcriptome Analysis of T3SS1 in Vibrio parahaemolyticus

Project description:Comparative transcriptional mRNA profiles were generated of bacterial pathogen Vibrio parahaemolyticus under conditions that induce activity of virulence factor type III secretion system 1 (T3SS1). Induction conditions included growth of the bacteria in Dulbecco's Modified Eagle Medium (DMEM), overexpression of transcriptional activator exsA and contact with HeLa cells in Hank's Balanced Salt Solution (HBSS), while non-inducing conditions included growth in Luria-Bertani medium supplemented with 2.5% (w/v) NaCl (LB-S) and overexpression of transcriptional anti-activator exsD. Transcriptome profiles of induction conditions were cross-compared against background non-inducing conditions and time points during HeLa cell infection (2 hr, 3 hr, 4 hr, 6 hr, 8 hr) were compared against pre-infection (0 hr) to identify genes important in T3SS1 activity and pathogenesis. Duplicate mRNA transcriptome profiles of V. parahaemolyticus grown in T3SS1 inducing (DMEM, exsA expression) and non-inducing (LB-S, exsD expression) conditions as well as during HeLa cell infection (0 hr, 2 hr, 3 hr, 4 hr, 6 hr, 8 hr) were generated by deep sequencing using an Illumina MiSeq

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Calcium is a global regulator in Vibrio parahaemolyticus

Project description:Here we probe the global response to calcium in the marine bacterium and pathogen Vibrio parahaemolyticus by using transcriptome, reporter and phenotypic analyses. Swarming gene expression and motility were enhanced by calcium. Calcium also stimulated expression and function of one of the organism’s two type three secretion systems (T3SS1 but not the T3SS2). Although low calcium is an inducing signal for the T3SS of many organisms, calcium stimulation of T3SS has not been reported before. EGTA was also a stimulus for T3SS1 expression and activity; however this appeared to be the consequence of iron rather than calcium chelation. LafK, a key regulator of swarming genes, was found necessary for calcium induction of T3SS1 gene transcription and cytotoxicity. Regulation of swarming and T3SS1 were additionally linked by a negative feedback loop propagated by ExsA. Overexpression of exsA was used to probe the extent of the T3SS1 regulon and verify its coincident induction by calcium and EGTA. The calcium transcriptome analysis revealed a calcium-repressed LysR-type transcriptional regulator; CalR was shown to repress swarming and T3SS1 gene expression. Thus in V. parahaemolyticus, calcium influences gene expression and behavior and seems a signal pertinent for surface colonization and virulence.

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