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Front Microbiol. 2019 Dec 3;10:2780. doi: 10.3389/fmicb.2019.02780. eCollection 2019.

A Broad Spectrum Protein Glycosylation System Influences Type II Protein Secretion and Associated Phenotypes in Vibrio cholerae.

Author information

1
Institute of Molecular Biosciences, University of Graz, Graz, Austria.
2
Centre for Ecological and Evolutionary Synthesis, Department of Biosciences, University of Oslo, Oslo, Norway.
3
Institute of Pathology, Medical University of Graz, Graz, Austria.
4
Omics Center Graz, BioTechMed-Graz, Graz, Austria.
5
Department of Molecular Microbiology, Washington University School of Medicine in St. Louis, St. Louis, MO, United States.
6
BioTechMed-Graz, Graz, Austria.
7
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.

Abstract

Protein secretion plays a crucial role for bacterial pathogens, exemplified by facultative human-pathogen Vibrio cholerae, which secretes various proteinaceous effectors at different stages of its lifecycle. Accordingly, the identification of factors impacting on protein secretion is important to understand the bacterial pathophysiology. PglLVc, a predicted oligosaccharyltransferase of V. cholerae, has been recently shown to exhibit O-glycosylation activity with relaxed glycan specificity in an engineered Escherichia coli system. By engineering V. cholerae strains to express a defined, undecaprenyl diphosphate-linked glycoform precursor, we confirmed functional O-linked protein glycosylation activity of PglLVc in V. cholerae. We demonstrate that PglLVc is required for the glycosylation of multiple V. cholerae proteins, including periplasmic chaperones such as DegP, that are required for efficient type II-dependent secretion. Moreover, defined deletion mutants and complementation strains provided first insights into the physiological role of O-linked protein glycosylation in V. cholerae. RbmD, a protein with structural similarities to PglLVc and other established oligosaccharyltransferases (OTases), was also included in this phenotypical characterization. Remarkably, presence or absence of PglLVc and RbmD impacts the secretion of proteins via the type II secretion system (T2SS). This is highlighted by altered cholera toxin (CT) secretion, chitin utilization and biofilm formation observed in ΔpglL Vc and ΔrbmD single or double mutants. This work thus establishes a unique connection between broad spectrum O-linked protein glycosylation and the efficacy of type II-dependent protein secretion critical to the pathogen's lifecycle.

KEYWORDS:

O-OTase; Vibrio cholerae; biofilm; chaperone; post-translational modification; virulence

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