LygA retention on the surface of Listeria monocytogenes via its interaction with wall teichoic acid modulates bacterial homeostasis and virulence.

Wall teichoic acid (WTA) is the abundant cell wall-associated glycopolymer in Gram-positive bacteria, playing crucial roles in surface proteins retention, bacterial homeostasis, and virulence. The WTA glycosylation of Listeria monocytogenes is essential for surface anchoring of virulence factors, whereas the nature and function of the noncovalent interactions between cell wall-associated proteins and WTA are less unknown. In this study, we found that galactosylated WTA (Gal-WTA) of serovar (SV) 4h L. monocytogenes plays a key role in modulating the novel glycine-tryptophan (GW) domain-containing autolysin protein LygA through direct interactions. Gal-deficient WTA of Lm XYSN (ΔgalT) showed a dramatic reduction of LygA on the cell surface. We demonstrated that LygA binds to Gal-WTA through the GW domains, and the binding affinity is associated with the number of GW motifs. Moreover, we confirmed the direct Gal-dependent binding of the GW protein Auto from the type I WTA strain, which has no interaction with rhamnosylated WTA, indicating that the complexity of both WTA and GW proteins affect the coordination patterns. Importantly, we revealed the crucial roles of LygA in facilitating bacterial homeostasis as well as crossing the intestinal and blood-brain barriers. Altogether, our findings suggest that both the glycosylation patterns of WTA and a fixed numbers of GW domains are closely associated with the retention of LygA on the cell surface, which promotes the pathogenesis of L. monocytogenes within the host.

Transmembrane Protein LMxysn_1693 of Serovar 4h Listeria monocytogenes Is Associated with Bile Salt Resistance and Intestinal Colonization.

Listeria monocytogenes (Lm) is a ubiquitous foodborne pathogen comprising of 14 serotypes, of which serovar 4h isolates belonging to hybrid sub-lineage Ⅱ exhibit hypervirulent features. LMxysn_1693 of serovar 4h Lm XYSN, a member of genomic island-7 (GI-7), is predicted to a membrane protein with unknown function, which is conserved in serovar 4h Listeria monocytogenes. Under bile salts stress, Lm XYSN strain lacking LMxysn_1693 (XYSN∆1693) exhibited a stationary phase growth defect as well as a reduction in biofilm formation and strikingly down-regulated bile-salts-resistant genes and virulent genes. Particularly, LMxysn_1693 protein plays a crucial role in Lm XYSN adhesion and invasion to intestinal epithelial cells, as well as colonization in the ileum of mice. Taken together, these findings indicate that the LMxysn_1693 gene encodes a component of the putative ABC transporter system, synthetically interacts with genes involved in bile resistance, biofilm formation and virulence, and thus contributes to Listeria monocytogenes survival within and outside the host.