In vivo repressed genes of Vibrio cholerae reveal inverse requirements of an H+ /Cl- transporter along the gastrointestinal passage

Proceedings of the National Academy of Sciences of the United States of America
Fatih CakarStefan Schild

Abstract

The facultative human pathogen Vibrio cholerae changes its transcriptional profile upon oral ingestion by the host to facilitate survival and colonization fitness. Here, we used a modified version of recombination-based in vivo expression technology to investigate gene silencing during the in vivo passage, which has been understudied. Using a murine model of cholera, we screened a V. cholerae transposon library composed of 10,000 randomly generated reporter fusions and identified 101 in vivo repressed (ivr) genes. Our data indicate that constitutive expression of ivr genes reduces colonization fitness, highlighting the necessity to down-regulate these genes in vivo. For example, the ivr gene clcA, encoding an H+/Cl- transporter, could be linked to the acid tolerance response against hydrochloric acid. In a chloride-dependent manner, ClcA facilitates survival under low pH (e.g., the stomach), but its presence becomes detrimental under alkaline conditions (e.g., lower gastrointestinal tract). This pH-dependent clcA expression is controlled by the LysR-type activator AphB, which acts in concert with AphA to initiate the virulence cascade in V. cholerae after oral ingestion. Thus, transcriptional networks dictating induction of vir...Continue Reading

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Citations

Apr 15, 2020·Infection and Immunity·D E KunkleJ E Bina
Dec 18, 2020·Molecular Microbiology·Nina PennetzdorferJoachim Reidl
Feb 14, 2020·Cell Host & Microbe·Bat-Erdene Jugder, Paula I Watnick
Oct 30, 2021·Microbial Biotechnology·Ralph BertramChristopher F Schuster

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Methods Mentioned

BETA
RNAseq
PCR
Assay
dissection

Software Mentioned

RIVET
StepOne
TRIVET

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