A dual-chain assembly pathway generates the high structural diversity of cell-wall polysaccharides in Lactococcus lactis

Loading...
Thumbnail Image
Files
PIIS002192582030750X.pdf(2.61 MB)
Published Version
mmc1.pdf(1.61 MB)
Supplementary Material
Date
2019
Authors
Theodorou, Ilias
Courtin, Pascal
Palussière, Simon
Kulakauskas, Saulius
Bidnenko, Elena
Péchoux, Christine
Fenaille, François
Penno, Christophe
Mahony, Jennifer
van Sinderen, Douwe
Journal Title
Journal ISSN
Volume Title
Publisher
American Society for Biochemistry and Molecular Biology Inc.
Research Projects
Organizational Units
Journal Issue
Abstract
In Lactococcus lactis, cell-wall polysaccharides (CWPSs) act as receptors for many bacteriophages, and their structural diversity among strains explains, at least partially, the narrow host range of these viral predators. Previous studies have reported that lactococcal CWPS consists of two distinct components, a variable chain exposed at the bacterial surface, named polysaccharide pellicle (PSP), and a more conserved rhamnan chain anchored to, and embedded inside, peptidoglycan. These two chains appear to be covalently linked to form a large heteropolysaccharide. The molecular machinery for biosynthesis of both components is encoded by a large gene cluster, named cwps. In this study, using a CRISPR/Cas-based method, we performed a mutational analysis of the cwps genes. MALDI-TOF MS-based structural analysis of the mutant CWPS combined with sequence homology, transmission EM, and phage sensitivity analyses enabled us to infer a role for each protein encoded by the cwps cluster. We propose a comprehensive CWPS biosynthesis scheme in which the rhamnan and PSP chains are independently synthesized from two distinct lipid-sugar precursors and are joined at the extracellular side of the cytoplasmic membrane by a mechanism involving a membrane-embedded glycosyltransferase with a GT-C fold. The proposed scheme encompasses a system that allows extracytoplasmic modification of rhamnan by complex substituting oligo-/polysaccharides. It accounts for the extensive diversity of CWPS structures observed among lactococci and may also have relevance to the biosynthesis of complex rhamnose-containing CWPSs in other Gram-positive bacteria.
Description
Keywords
Gram-positive bacteria , Polysaccharide , Cell wall , Biosynthesis , Bacteriophage , Glycopolymer , Glycosyltransferase , Lactic acid bacteria , Rhamnan , Rhamnose
Citation
Theodorou, I., Courtin, P., Palussière, S., Kulakauskas, S., Bidnenko, E., Péchoux, C., Fenaille, F., Penno, C., Mahony, J., Van Sinderen, D. and Chapot-Chartier, M.-P. (2019) ‘A dual-chain assembly pathway generates the high structural diversity of cell-wall polysaccharides in Lactococcus lactis’, Journal of Biological Chemistry, 294(46), pp. 17612–17625. doi: 10.1074/jbc.RA119.009957
Link to publisher’s version