Role of glycosyltransferases modifying type B flagellin of emerging hypervirulent Clostridium difficile lineages and their impact on motility and biofilm formation
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
Clostridium difficile is the principal cause of nosocomial infectious diarrhea worldwide. The pathogen modifies its flagellin with either a type A or type B O-linked glycosylation system, which has a contributory role in pathogenesis. We study the functional role of glycosyltransferases modifying type B flagellin in the 023 and 027 hypervirulent C. difficile lineages by mutagenesis of five putative glycosyltransferases and biosynthetic genes. We reveal their roles in the biosynthesis of the flagellin glycan chain and demonstrate that flagellar post-translational modification affects motility and adhesion-related bacterial properties of these strains. We show that the glycosyltransferases 1 and 2 (GT1 and GT2) are responsible for the sequential addition of a GlcNAc and two rhamnoses, respectively, and that GT3 is associated with the incorporation of a novel sulfonated peptidyl-amido sugar moiety whose structure is reported in our accompanying paper (Bouché, L., Panico, M., Hitchen, P., Binet, D., Sastre, F., Faulds-Pain, A., Valiente, E., Vinogradov, E., Aubry, A., Fulton, K., Twine, S., Logan, S. M., Wren, B. W., Dell, A., and Morris, H. R. (2016) J. Biol. Chem. 291, 25439–25449). GT2 is also responsible for methylation of the rhamnoses. Whereas type B modification is not required for flagellar assembly, some mutations that result in truncation or abolition of the glycan reduce bacterial motility and promote autoaggregation and biofilm formation. The complete lack of flagellin modification also significantly reduces adhesion of C. difficile to Caco-2 intestinal epithelial cells but does not affect activation of human TLR5. Our study advances our understanding of the genes involved in flagellar glycosylation and their biological roles in emerging hypervirulent C. difficile strains.
Date Issued
2016-10-04
Date Acceptance
2016-09-27
Citation
Journal of Biological Chemistry, 2016, 291 (49), pp.25450-25461
ISSN
1083-351X
Publisher
American Society for Biochemistry and Molecular Biology
Start Page
25450
End Page
25461
Journal / Book Title
Journal of Biological Chemistry
Volume
291
Issue
49
Copyright Statement
© 2016 by The American Society for Biochemistry and Molecular Biology, Inc. Final version free via Creative Commons CC-BY license (https://creativecommons.org/licenses/by/4.0/).
Sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
Wellcome Trust
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000390095300015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Grant Number
BB/K016164/1
102978/Z/13/Z
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
bacteria
biofilm
glycosylation
glycosyltransferase
mutant
Clostridium difficile
flagella
gram positive bacteria
CAMPYLOBACTER-JEJUNI
GLYCOSYLATION ISLAND
ESCHERICHIA-COLI
TOXIN-B
PROTEIN
COLONIZATION
INFECTION
BIOSYNTHESIS
EPIDEMIOLOGY
VIRULENCE
06 Biological Sciences
11 Medical And Health Sciences
03 Chemical Sciences
Publication Status
Published