The tip protein PAAR is required for the function of the type VI secretion system

Solène G Beauvois (1) , Nicolas Flaugnatti (1) , Marianne Ilbert (2) , Marie Boyer (1) , Esther Gavello-Fernandez (3, 4) , Rémi Fronzes (3, 4) , Dukas Jurėnas (1) , Laure Journet (1)

1 LISM - Laboratoire d'ingénierie des systèmes macromoléculaires
2 BIP - Bioénergétique et Ingénierie des Protéines
3 IECB - Institut Européen de Chimie et Biologie
4 MFP - Microbiologie Fondamentale et Pathogénicité

Bacteria are constantly competing to colonize crowded ecological niches, such as the human gut. The type VI secretion system (T6SS) is a critical bacterial weapon in this warfare. It resembles a crossbow with a poisoned arrow allowing bacteria to inject toxic effectors directly into target cells. This machinery is formed by an envelopespanning complex which recruits the baseplate, an assembly platform allowing the polymerization of a contractile structure. The tail consists of a tube surrounded by a sheath and topped by the needle complex composed of the VgrG and PAAR proteins. In the enteric pathogen enteroaggregative Escherichia coli (EAEC), the Tle1 phospholipase toxin ensures the antibacterial activity of the T6SS-1. For its transport, Tle1 interacts directly with the trimeric spike protein VgrG. However, the importance and the function of the tip protein PAAR in the T6SS remain unclear. Here, we characterized the PAAR protein of EAEC using biochemical, fluorescence microscopy, and antibacterial competi tion approaches. Using pull-down assays and cryo-electron microscopy analysis of the (VgrG) 3-(Tle1) 3-PAAR complex, we show that PAAR tops and closes the β-prism structure of the VgrG spike. The PAAR protein structure is further tightened by the zinc atom coordinated via conserved residues essential for its function. We provide evidence that PAAR is necessary for T6SS-1-mediated killing due to its requirement for proper T6SS baseplate assembly and further sheath polymerization. Our results suggest that the PAAR protein is an essential component of T6SS.

Mots clés: Type VI secretion system; cryo-electron microscopy; bacterial competition; protein secretion; VgrG; bacterial toxin; zinc

Domaines: Biochimie, Biologie Moléculaire

Solène G Beauvois, Nicolas Flaugnatti, Marianne Ilbert, Marie Boyer, Esther Gavello-Fernandez, et al.. The tip protein PAAR is required for the function of the type VI secretion system. Microbiology Spectrum, 2023, ⟨10.1128/spectrum.01478-23⟩. ⟨hal-04260438⟩