Oral Presentation Australian Society for Microbiology Annual Scientific Meeting 2022

Deployment of the type six secretion system arsenal by a sigma factor and enhancer binding protein in Pseudomonas aeruginosa (82046)

Luke P Allsopp 1 , Alice CZ Collins 1 , Elanor Hawkins 2 , Thomas E Wood 2 , Alain Filloux 2
  1. National Heart and Lung Institute, Imperial College London, London, United Kingdom
  2. Department of Life Sciences, MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, United Kingdom

The type VI secretion system (T6SS) is a molecular nanomachine which contributes to infection by injecting potent toxins via a syringe-like complex directly into host cells or competing microorganisms. Pseudomonas aeruginosa encodes three distinct T6SS machines (H1-, H2-, H3-) in separate gene clusters, which are mainly associated with Gram-negative bacterial competition. Additionally, multiple orphan gene clusters are spread throughout the genome that encode different T6SS VgrG puncturing tips and known or putative toxic effector proteins. However, the deployment of each system and mechanism of engagement of orphan gene clusters to facilitate expression and loading of a wide variety of toxic effector payloads is unknown. As the major alternative sigma factor RpoN is associated with flagella, virulence factors and cell surface control, we hypothesised that RpoN may facilitate T6SS expression control in P. aeruginosa. Using RNAseq, ChIPseq and molecular biology approaches we demonstrate that RpoN coordinates the T6SSs of P. aeruginosa, activating the H2-T6SS but repressing the H1- and H3-T6SS. Further studies showed the combined action of RpoN with a designated sigma factor activator protein Sfa2 enabled coordination of tip and effector expression to enable interbacterial killing via the H2-T6SS. Furthermore, the posttranscriptional regulator RsmA showed global coordinated repression of almost all T6SS genes. Our study further delineates the multiple regulatory mechanisms that modulate the deployment of an arsenal of T6SS effectors. This enables a versatile organism like P. aeruginosa to respond and adapt to a range of environmental conditions and competitive settings allowing it to thrive.

  1. Allsopp LP†, Collins ACZ, Hawkins E, Wood TE, Filloux A†. (2022) RpoN/Sfa2-dependent activation of the Pseudomonas aeruginosa H2-T6SS and its cognate arsenal of antibacterial toxins. Nucleic Acid Research. doi:10.1093/nar/gkab1254
  2. Allsopp LP†, Bernal P, Nolan LM, Filloux A†. (2020) Causalities of War: The connection between T6SS and microbiota. Cell Microbiol. doi: 10.1111/cmi.13153
  3. Allsopp LP, Wood TE, Howard SA, Maggiorelli F, Nolan LM, Wettstadt S, Filloux A. (2017) RsmA and AmrZ orchestrate the assembly of all three type VI secretion systems in Pseudomonas aeruginosa. PNAS. doi: 10.1073/pnas.1700286114