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The post‐translational modification of the C lostridium difficile flagellin affects motility, cell surface properties and virulence
Author(s) -
FauldsPain Alexandra,
Twine Susan M.,
Vinogradov Evgeny,
Strong Philippa C. R.,
Dell Anne,
Buckley Anthony M.,
Douce Gillian R.,
Valiente Esmeralda,
Logan Susan M.,
Wren Brendan W.
Publication year - 2014
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.12755
Subject(s) - flagellin , biology , threonine , glycan , motility , microbiology and biotechnology , mutant , virulence , serine , open reading frame , biochemistry , phosphorylation , gene , peptide sequence , glycoprotein
Summary C lostridium 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 CD 0241 encodes a kinase involved in the transfer of the phosphate to the threonine, the CD 0242 protein catalyses the addition of the phosphothreonine to the N ‐acetylglucosamine moiety and CD 0243 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.