# Morphogenetic penicillin-binding proteins control virulence-associated type III secretion systems in Salmonella

## TL;DR

This study shows how specific penicillin-binding proteins in Salmonella influence the activity of virulence-related protein secretion systems.

## Contribution

The paper provides the first evidence linking morphogenetic penicillin-binding proteins to virulence-associated T3SS in Salmonella.

## Key findings

- PBP2 is required for proper function of the SPI-1 T3SS but not for motility.
- Lack of morphogenetic PBPs increases SPI-2 T3SS activity.
- PBP2 controls SPI-1 T3SS via regulators HilA and InvF.

## Abstract

Type III protein secretion systems (T3SSs) function as multiprotein devices that span the envelope of Gram-negative bacteria using the peptidoglycan (PG) layer as scaffold. This spatial arrangement explains why modifications in PG structure can alter T3SS activity. In Salmonella, incorporation of non-canonical D-amino acids in the PG was shown to decrease the activity of the T3SS encoded by the pathogenicity island-1 (SPI-1) without affecting other T3SS, like the flagellum apparatus. Enigmatically, following invasion of host cell Salmonella enterica serovar Typhimurium modifies PG synthesis by upregulating two pathogen-specific enzymes, the penicillin-binding proteins PBP2SAL and PBP3SAL, with roles in cell elongation and division, respectively. In the mouse typhoid model, the amount of PBP2SAL and PBP3SAL produced by the pathogen exceeds by large those of the canonical enzymes PBP2 and PBP3. This change responds to acidity and high osmolarity, the same cues that intra-phagosomal S. Typhimurium perceives to switch the SPI-1 T3SS by that encoded in SPI-2. Using isogenic mutants lacking each of the four morphogenetic PBPs, we tested whether their activities and those of the T3SS encoded by SPI-1 and SPI-2, are interconnected. Our data show that PBP2 is required for proper function of SPI-1 T3SS but dispensable for motility, whereas the lack of any of the morphogenetic PBPs increases SPI-2 T3SS activity. The positive control exerted by PBP2 on SPI-1 takes place via the SPI-1-specific regulators HilA and InvF. To our knowledge, these findings provide the first evidence linking morphogenetic enzymes that synthesize PG with T3SS associated to virulence.