Nov 8, 2018

New players in the regulatory network of Vibrio parahaemolyticus type VI secretion system 1

BioRxiv : the Preprint Server for Biology
Rotem Ben-Yaakov, Dor Salomon


Type VI secretion systems (T6SSs) are widespread, tightly regulated, protein delivery apparatuses used by Gram-negative bacteria to outcompete their neighbors. The pathogen, Vibrio parahaemolyticus , encodes two T6SSs. These T6SSs are differentially regulated by external conditions. T6SS1, an antibacterial system predominantly found in pathogenic isolates, requires warm marine-like conditions and surface sensing for activation. The regulatory network that governs this activation is not well understood. In this work, we devised a screening methodology that allows us to easily monitor the outcome of bacterial competitions and thus, to identify mutants that are defective in T6SS1-mediated bacterial killing. The methodology, termed Bacterial Competition Fluorescence (BaCoF), relies on detection of a fluorescent signal as an indicator of the survival and growth of a T6SS-sensitive, GFP-expressing prey that has been co-cultured with mutants derived from a T6SS+ attacker of interest. Using BaCoF, we screened a random transposon insertion mutant library and identified genes required for V. parahaemolyticus T6SS1 activation, among them TfoY and Tmk. We used epistasis experiments to determine the relationships between the newly identifie...Continue Reading

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Mentioned in this Paper

Fluorescent Dyes
Protein Secretion by the Type VI Secretion System
Vibrio parahaemolyticus
Regulation of Biological Process
Pathogenic Organism
Coculture Techniques
Mutant Proteins

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