The bacterial toxin ExoU requires a host trafficking chaperone for transportation and to induce necrosis

Vincent Deruelle,Ina Attrée,Marie-Odile Fauvarque,Philippe Huber,Viviana Job,Emmanuel Taillebourg,Stéphanie Bouillot

doi:10.1038/s41467-021-24337-9

Abstract

Pseudomonas aeruginosa can cause nosocomial infections, especially in ventilated or cystic fibrosis patients. Highly pathogenic isolates express the phospholipase ExoU, an effector of the type III secretion system that acts on plasma membrane lipids, causing membrane rupture and host cell necrosis. Here, we use a genome-wide screen to discover that ExoU requires DNAJC5, a host chaperone, for its necrotic activity. DNAJC5 is known to participate in an unconventional secretory pathway for misfolded proteins involving anterograde vesicular trafficking. We show that DNAJC5-deficient human cells, or Drosophila flies knocked-down for the DNAJC5 orthologue, are largely resistant to ExoU-dependent virulence. ExoU colocalizes with DNAJC5-positive vesicles in the host cytoplasm. DNAJC5 mutations preventing vesicle trafficking (previously identified in adult neuronal ceroid lipofuscinosis, a human congenital disease) inhibit ExoU-dependent cell lysis. Our results suggest that, once injected into the host cytoplasm, ExoU docks to DNAJC5-positive secretory vesicles to reach the plasma membrane, where it can exert its phospholipase activity

Highlights

Pseudomonas aeruginosa can cause nosocomial infections, especially in ventilated or cystic fibrosis patients
The cells were subjected to three rounds of infection with the P. aeruginosa strain PA14, known to induce cell necrosis via ExoU secretion (Fig. 1a)
Three independent replicates were performed and a statistical analysis revealed a significant enrichment for gRNAs targeting only one gene: the gene encoding DNAJC5(Fig. 1b)

Summary

Introduction

Pseudomonas aeruginosa can cause nosocomial infections, especially in ventilated or cystic fibrosis patients. Once injected into the host cytoplasm, ExoU docks to DNAJC5-positive secretory vesicles to reach the plasma membrane, where it can exert its phospholipase activity. The requirement for host-cell mechanisms protects bacteria from self-toxicity and takes advantage of efficient molecular mechanisms developed by eukaryotic cells to alter cellular functions This rule applies to the toxins secreted by Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen. P. aeruginosa uses a multi-target strategy to infect host cells, employing a combination of virulence factors. One of these factors is the type 3 secretion system (T3SS), the effectors of which are known to be the most potent toxins in acute P. aeruginosa infections[2,7]. ExoU-positive strains have been associated with increased multidrug resistance in several clinical studies[16,17,18,19]

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