AUTHOR=Zhang Yani , Zhang Chao , Du Xiao , Zhou Yun , Kong Weina , Lau Gee W. , Chen Gukui , Kohli Gurjeet Singh , Yang Liang , Wang Tietao , Liang Haihua 

TITLE=Glutathione Activates Type III Secretion System Through Vfr in Pseudomonas aeruginosa

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=9

YEAR=2019

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2019.00164

DOI=10.3389/fcimb.2019.00164

ISSN=2235-2988

ABSTRACT=<p>Glutathione (GSH) is the most abundant antioxidant in all living organisms. Previously, we have shown that a deletion mutant in the glutathione synthetase gene (Δ<italic>gshB</italic>) decreases the expression of type III secretion system (T3SS) genes of <italic>Pseudomonas aeruginosa</italic>. However, the mechanism remains elusive. In this study, a comprehensive transcriptomic analysis of the GSH-deficient mutant Δ<italic>gshA</italic>Δ<italic>gshB</italic> was used to elucidate the role of GSH in the pathogenesis of <italic>P. aeruginosa</italic>. The data show that the expression of genes in T3SS, type VI secretion system (T6SS) and some regulatory genes were impaired. Δ<italic>gshA</italic>Δ<italic>gshB</italic> was attenuated in a mouse model of acute pneumonia, swimming and swarming motilities, and biofilm formation. Under T3SS inducing conditions, GSH enhanced the expression of T3SS in both wild-type PAO1 and Δ<italic>gshA</italic>Δ<italic>gshB</italic>, but not in Δ<italic>vfr</italic>. Genetic complementation of Δ<italic>vfr</italic> restored the ability of GSH to induce the expression of T3SS genes. Site-directed mutagenesis based substitution of cysteine residues with alanine in Vfr protein abolished the induction of T3SS genes by GSH, confirming that GSH regulates T3SS genes through Vfr. Exposure to H<sub>2</sub>O<sub>2</sub> decreased free thiol content on Vfr, indicating that the protein was sensitive to redox modification. Importantly, GSH restored the oxidized Vfr to reduced state. Collectively, these results suggest that GSH serves as an intracellular redox signal sensed by Vfr to upregulate T3SS expression in <italic>P. aeruginos</italic>a. Our work provides new insights into the role of GSH in <italic>P. aeruginosa</italic> pathogenesis.</p>