The post-translational modification of theClostridium difficileflagellin affects motility, cell surface properties and virulence

Faulds-Pain, A., Twine, S. M., Vinogradov, E., Strong, P. C. R., Dell, A., Buckley, A. M., Douce, G. R. , Valiente, E., Logan, S. M. and Wren, B. W. (2014) The post-translational modification of theClostridium difficileflagellin affects motility, cell surface properties and virulence. Molecular Microbiology, 94(2), pp. 272-289. (doi: 10.1111/mmi.12755) (PMID:25135277) (PMCID:PMC4441256)

[img]
Preview
Text
106027.pdf - Published Version
Available under License Creative Commons Attribution.

3MB

Abstract

Clostridium difficile is a prominent nosocomial pathogen, proliferating and causing enteric disease in individuals with a compromised gut microflora. We characterized the post-translational modification of flagellin in C. difficile 630. The structure of the modification was solved by nuclear magnetic resonance and shown to contain an N-acetylglucosamine substituted with a phosphorylated N-methyl-l-threonine. A reverse genetics approach investigated the function of the putative four-gene modification locus. All mutants were found to have truncated glycan structures by LC-MS/MS, taking into account bioinformatic analysis, we propose that the open reading frame CD0241 encodes a kinase involved in the transfer of the phosphate to the threonine, the CD0242 protein catalyses the addition of the phosphothreonine to the N-acetylglucosamine moiety and CD0243 transfers the methyl group to the threonine. Some mutations affected motility and caused cells to aggregate to each other and abiotic surfaces. Altering the structure of the flagellin modification impacted on colonization and disease recurrence in a murine model of infection, showing that alterations in the surface architecture of C. difficile vegetative cells can play a significant role in disease. We show that motility is not a requirement for colonization, but that colonization was compromised when the glycan structure was incomplete.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Douce, Dr Gillian and Buckley, Dr Anthony
Authors: Faulds-Pain, A., Twine, S. M., Vinogradov, E., Strong, P. C. R., Dell, A., Buckley, A. M., Douce, G. R., Valiente, E., Logan, S. M., and Wren, B. W.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Molecular Microbiology
Publisher:John Wiley & Sons Ltd.
ISSN:0950-382X
ISSN (Online):1365-2958
Copyright Holders:Copyright © 2014 The Authors
First Published:First published in Molecular Microbiology 94(2):272-289
Publisher Policy:Reproduced under a Creative Commons License

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
492111Genetic and phenotypic characterisation of emerging virulent Clostridium difficileGillian DouceWellcome Trust (WELLCOME)086418/B/08/ZIII - BACTERIOLOGY