Critical role for a promoter discriminator in RpoS control of virulence in Edwardsiella piscicida

PLoS Pathogens
Kaiyu YinQiyao Wang

Abstract

Edwardsiella piscicida is a leading fish pathogen that causes significant economic loses in the aquaculture industry. The pathogen depends on type III and type VI secretion systems (T3/T6SS) for growth and virulence in fish and the expression of both systems is controlled by the EsrB transcription activator. Here, we performed a Tn-seq-based screen to uncover factors that govern esrB expression. Unexpectedly, we discovered that RpoS antagonizes esrB expression and thereby inhibits production of E. piscicida's T3/T6SS. Using in vitro transcription assays, we showed that RpoS can block RpoD-mediated transcription of esrB. ChIP-seq- and RNA-seq-based profiling, as well as mutational and biochemical analyses revealed that RpoS-repressed promoters contain a -6G in their respective discriminator sequences; moreover, this -6G proved critical for RpoS to inhibit esrB expression. Mutation of the RpoS R99 residue, an amino acid that molecular modeling predicts interacts with -6G in the esrB discriminator, abolished RpoS' capacity for repression. In a turbot model, an rpoS deletion mutant was attenuated early but not late in infection, whereas a mutant expressing RpoSR99A exhibited elevated fitness throughout the infection period. Collect...Continue Reading

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Citations

May 28, 2019·Virulence·Ka Yin LeungBupe A Siame
Mar 17, 2019·Applied and Environmental Microbiology·Lifan WeiQiyao Wang
Oct 13, 2019·IScience·Lifan WeiQiyao Wang

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Methods Mentioned

BETA
dissection
RNA-seq
environmental stresses
pull-down
ChIP
In vitro transcription
PCR
Fluorescence
immunoprecipitation assay
ChIP-seq

Software Mentioned

Trimmomatic
Con
ARTIST
ChIP
MEME
GENEWIZ

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