Glutathione activates type III secretion system through Vfr in pseudomonas aeruginosa

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

Research output: Contribution to journalArticle

Abstract

Glutathione (GSH) is the most abundant antioxidant in all living organisms. Previously, we have shown that a deletion mutant in the glutathione synthetase gene (ΔgshB) decreases the expression of type III secretion system (T3SS) genes of Pseudomonas aeruginosa. However, the mechanism remains elusive. In this study, a comprehensive transcriptomic analysis of the GSH-deficient mutant ΔgshAΔgshB was used to elucidate the role of GSH in the pathogenesis of P. aeruginosa. The data show that the expression of genes in T3SS, type VI secretion system (T6SS) and some regulatory genes were impaired. ΔgshAΔgshB 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 ΔgshAΔgshB, but not in Δvfr. Genetic complementation of Δvfr 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 H2O2 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 P. aeruginosa. Our work provides new insights into the role of GSH in P. aeruginosa pathogenesis.

Original languageEnglish (US)
Article number164
JournalFrontiers in Cellular and Infection Microbiology
Volume9
Issue numberMAY
DOIs
StatePublished - Jan 1 2019

Fingerprint

Pseudomonas aeruginosa
Glutathione
Genes
Oxidation-Reduction
Glutathione Synthase
Gene Expression
Regulator Genes
Biofilms
Site-Directed Mutagenesis
Sulfhydryl Compounds
Alanine
Cysteine
Pneumonia
Proteins
Up-Regulation
Antioxidants
Type III Secretion Systems

Keywords

  • Glutathione
  • Pathogenicity
  • Pseudomonas aeruginosa
  • Type III secretion system
  • Vfr

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

Cite this

Glutathione activates type III secretion system through Vfr in pseudomonas aeruginosa. / Zhang, Yani; Zhang, Chao; Du, Xiao; Zhou, Yun; Kong, Weina; Lau, Gee; Chen, Gukui; Kohli, Gurjeet Singh; Yang, Liang; Wang, Tietao; Liang, Haihua.

In: Frontiers in Cellular and Infection Microbiology, Vol. 9, No. MAY, 164, 01.01.2019.

Research output: Contribution to journalArticle

Zhang, Y, Zhang, C, Du, X, Zhou, Y, Kong, W, Lau, G, Chen, G, Kohli, GS, Yang, L, Wang, T & Liang, H 2019, 'Glutathione activates type III secretion system through Vfr in pseudomonas aeruginosa', Frontiers in Cellular and Infection Microbiology, vol. 9, no. MAY, 164. https://doi.org/10.3389/fcimb.2019.00164
Zhang, Yani ; Zhang, Chao ; Du, Xiao ; Zhou, Yun ; Kong, Weina ; Lau, Gee ; Chen, Gukui ; Kohli, Gurjeet Singh ; Yang, Liang ; Wang, Tietao ; Liang, Haihua. / Glutathione activates type III secretion system through Vfr in pseudomonas aeruginosa. In: Frontiers in Cellular and Infection Microbiology. 2019 ; Vol. 9, No. MAY.
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AU - Zhang, Chao

AU - Du, Xiao

AU - Zhou, Yun

AU - Kong, Weina

AU - Lau, Gee

AU - Chen, Gukui

AU - Kohli, Gurjeet Singh

AU - Yang, Liang

AU - Wang, Tietao

AU - Liang, Haihua

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AB - Glutathione (GSH) is the most abundant antioxidant in all living organisms. Previously, we have shown that a deletion mutant in the glutathione synthetase gene (ΔgshB) decreases the expression of type III secretion system (T3SS) genes of Pseudomonas aeruginosa. However, the mechanism remains elusive. In this study, a comprehensive transcriptomic analysis of the GSH-deficient mutant ΔgshAΔgshB was used to elucidate the role of GSH in the pathogenesis of P. aeruginosa. The data show that the expression of genes in T3SS, type VI secretion system (T6SS) and some regulatory genes were impaired. ΔgshAΔgshB 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 ΔgshAΔgshB, but not in Δvfr. Genetic complementation of Δvfr 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 H2O2 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 P. aeruginosa. Our work provides new insights into the role of GSH in P. aeruginosa pathogenesis.

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