Type III Secretion Effector VopQ of Vibrio parahaemolyticus Modulates Central Carbon Metabolism in Epithelial Cells.

Anh Quoc Nguyen,Akira Takahashi,Sho Hatayama,Junko Kido,Aya Tentaku,Takaaki Shimohata,Thi Mai Huong Bui,Takashi Uebanso,Kazuaki Mawatari,Sarah E.F D'Orazio

doi:10.1128/msphere.00960-19

Abstract

Vibrio parahaemolyticus is a Gram-negative halophilic pathogen that frequently causes acute gastroenteritis and occasional wound infection. V. parahaemolyticus contains several virulence factors, including type III secretion systems (T3SSs) and thermostable direct hemolysin (TDH). In particular, T3SS1 is a potent cytotoxic inducer, and T3SS2 is essential for causing acute gastroenteritis. Although much is known about manipulation of host signaling transductions by the V. parahaemolyticus effector, little is known about the host metabolomic changes modulated by V. parahaemolyticus To address this knowledge gap, we performed a metabolomic analysis of the epithelial cells during V. parahaemolyticus infection using capillary electrophoresis-time of flight mass spectrometry (CE-TOF/MS). Our results revealed significant metabolomic perturbations upon V. parahaemolyticus infection. Moreover, we identified that T3SS1's VopQ effector was responsible for inducing the significant metabolic changes in the infected cells. The VopQ effector dramatically altered the host cell's glycolytic, tricarboxylic acid cycle (TCA), and amino acid metabolisms. VopQ effector disrupted host cell redox homeostasis by depleting cellular glutathione and subsequently increasing the level of reactive oxygen species (ROS) production.IMPORTANCE The metabolic response of host cells upon infection is pathogen specific, and infection-induced host metabolic reprogramming may have beneficial effects on the proliferation of pathogens. V. parahaemolyticus contains a range of virulence factors to manipulate host signaling pathways and metabolic processes. In this study, we identified that the T3SS1 VopQ effector rewrites host metabolism in conjunction with the inflammation and cell death processes. Understanding how VopQ reprograms host cell metabolism during the infection could help us to identify novel therapeutic strategies to enhance the survival of host cells during V. parahaemolyticus infection.

Highlights

Vibrio parahaemolyticus is a Gram-negative halophilic pathogen that frequently causes acute gastroenteritis and occasional wound infection
We investigated the dynamics of the metabolic processes of the epithelial cells during V. parahaemolyticus infection and the role of the VopQ effector of T3SS1 in modulating host cell metabolism using capillary electrophoresis-time of flight mass spectrometry (CE-TOF/MS)
The levels of subsequent downstream glycolytic intermediates were altered in Caco-2 cells infected with ET4 or POR3 compared to S1-ENM and the control. In those infected with strains ET4 and POR3, we found significant reductions of downstream glycolytic intermediates, including fructose 1,6-phosphate (F-1,6-P), 3-phosphoglycerate (3GP), phospho(enol)pyruvic acid (PEP), and pyruvic acid (PYR) (P Ͻ 0.05) (Fig. 3), compared to Caco-2 cells infected with the S1-ENM mutant or with the control

Summary

Introduction

Vibrio parahaemolyticus is a Gram-negative halophilic pathogen that frequently causes acute gastroenteritis and occasional wound infection. Much is known about manipulation of host signaling transductions by the V. parahaemolyticus effector, little is known about the host metabolomic changes modulated by V. parahaemolyticus To address this knowledge gap, we performed a metabolomic analysis of the epithelial cells during V. parahaemolyticus infection using capillary electrophoresis-time of flight mass spectrometry (CE-TOF/MS). Clinical isolates of V. parahaemolyticus contain numerous virulence factors, including pore-forming thermostable direct hemolysin (TDH) toxin and two type III secretion systems (T3SSs) that enable the delivery of bacterial effectors into the eukaryotic host [5, 6]. We investigated the dynamics of the metabolic processes of the epithelial cells during V. parahaemolyticus infection and the role of the VopQ effector of T3SS1 in modulating host cell metabolism using capillary electrophoresis-time of flight mass spectrometry (CE-TOF/MS). We identified that the cytotoxic VopQ effector of T3SS1 is a metabolic disruptor, profoundly altering host metabolisms

Methods

Results

Conclusion