Discrete and overlapping functions of peptidoglycan synthases in growth, cell division and virulence of Listeria monocytogenes

• DOI: 10.1111/mmi.12873
• Publication Date: 2014-11-26T02:27:49Z
• Authors: Jeanine Rismondo; Lars Möller; Christine Aldridge; Joe Gray; Waldemar Vollmer; Sven Halbedel
• Keywords: 0301 basic medicine; Penicillin-Binding Proteins/genetics; 610 Medizin; 610; Microbial Sensitivity Tests; Peptidoglycan; beta-Lactams; beta-Lactams/pharmacology; Anti-Bacterial Agents/pharmacology; Virulence/genetics; Peptidoglycan/metabolism; Mice; 03 medical and health sciences; Bacterial Proteins; Cell Wall; Listeria monocytogenes/cytology; Animals; Humans; Penicillin-Binding Proteins; Research Articles; ddc:610; Virulence; Bacterial Proteins/metabolism; Cell Wall/physiology; 3T3 Cells; Cell Wall/chemistry; Listeria monocytogenes; Penicillin-Binding Proteins/metabolism; Anti-Bacterial Agents; Listeria monocytogenes/pathogenicity; Listeria monocytogenes/physiology; Mutation; Listeria monocytogenes/drug effects; HeLa Cells

SummaryUpon ingestion of contaminated food, Listeria monocytogenes can cause serious infections in humans that are normally treated with β‐lactam antibiotics. These target Listeria's five high molecular weight penicillin‐binding proteins (HMW PBPs), which are required for peptidoglycan biosynthesis. The two bi‐functional class A HMW PBPs PBP A1 and PBP A2 have transglycosylase and transpeptidase domains catalyzing glycan chain polymerization and peptide cross‐linking, respectively, whereas the three class B HMW PBPs B1, B2 and B3 are monofunctional transpeptidases. The precise roles of these PBPs in the cell cycle are unknown. Here we show that green fluorescent protein (GFP)‐PBP fusions localized either at the septum, the lateral wall or both, suggesting distinct and overlapping functions. Genetic data confirmed this view: PBP A1 and PBP A2 could not be inactivated simultaneously, and a conditional double mutant strain is largely inducer dependent. PBP B1 is required for rod‐shape and PBP B2 for cross‐wall biosynthesis and viability, whereas PBP B3 is dispensable for growth and cell division. PBP B1 depletion dramatically increased β‐lactam susceptibilities and stimulated spontaneous autolysis but had no effect on peptidoglycan cross‐linkage. Our in vitro virulence assays indicated that the complete set of all HMW PBPs is required for maximal virulence.