Type

Journal Article

Authors

Douwe van Sinderen
Marie-Pierre Chapot-Chartier
Jennifer Mahony
François Fenaille
Simon Palussière
Irina Sadovskaya
Pascal Courtin
Ilias Theodorou

Subjects

Microbiology

Topics
lipoteichoic acid cell wall phage receptor glycosyltransferase glycopolymer lta flippase genomics bacteriophage lactic acid bacteria glycobiology peptidoglycan glycosylation

Three distinct glycosylation pathways are involved in the decoration of Lactococcus lactis cell wall glycopolymers (2020)

Abstract Extra-cytoplasmic sugar decoration of glycopolymer components of the bacterial cell wall contributes to their structural diversity. Typically, the molecular mechanism that underpins such a decoration process involves a three-component glycosylation system (TGS) represented by an undecaprenyl-phosphate (Und-P) sugar-activating glycosyltransferase (Und-P GT), a flippase, and a polytopic glycosyltransferase (PolM GT) dedicated to attaching sugar residues to a specific glycopolymer. Here, using bioinformatic analyses, CRISPR-assisted recombineering, structural analysis of cell wall-associated polysaccharides (CWPS) through Maldi-Tof MS and methylation analysis, we report on three such systems in the bacterium Lactococcus lactis. On the basis of sequence similarities, we first identified three gene pairs, csdAB, csdCD, and csdEF, each encoding an Und-P GT and a PolM GT, as potential TGS component candidates. Our experimental results show that csdAB and csdCD are involved in Glc side chain addition on the CWPS components rhamnan and polysaccharide pellicle (PSP), respectively, whereas csdEF plays a role in galactosylation of lipoteichoic acid (LTA). We also identified a potential flippase encoded in the L. lactis genome (llnz_02975, cflA) and confirmed that it participates in the glycosylation of the three cell wall glycopolymers rhamnan, PSP, and LTA, thus indicating that its function is shared by the three TGSs. Finally, we observed that glucosylation of both rhamnan and PSP can increase resistance to bacteriophage predation and that LTA galactosylation alters L. lactis resistance to bacteriocin.
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Full list of authors on original publication

Douwe van Sinderen, Marie-Pierre Chapot-Chartier, Jennifer Mahony, François Fenaille, Simon Palussière, Irina Sadovskaya, Pascal Courtin, Ilias Theodorou

Experts in our system

1
Douwe van Sinderen
University College Cork
Total Publications: 144
 
2
Marie-Pierre Chapot-Chartier
University College Cork
 
3
Jennifer Mahony
University College Cork
Total Publications: 87
 
4
Irina Sadovskaya
University College Cork
Total Publications: 10
 
5
Pascal Courtin
University College Cork