FLAUGNATTI, Nicolas; RAPISARDA, Chiara; REY, Martial; BEAUVOIS, Solène; NGUYEN, Viet Anh; CANAAN, Stéphane; DURAND, Eric; CHAMOT‐ROOKE, Julia; CASCALES, E.; FRONZES, Rémi; JOURNET, Laure

doi:10.15252/embj.2019104129

RAPISARDA, Chiara
Microbiologie Fondamentale et Pathogénicité [MFP]

REY, Martial

Langue

Article de revue

Ce document a été publié dans

EMBO Journal. 2020-06-02, vol. 39, n° 11, p. e104129

Résumé en anglais

The bacterial type VI secretion system (T6SS) is a macromolecular machine that injects effectors into prokaryotic and eukaryotic cells. The mode of action of the T6SS is similar to contractile phages: the contraction of a sheath structure pushes a tube topped by a spike into target cells. Effectors are loaded onto the spike or confined into the tube. In enteroaggregative Escherichia coli, the Tle1 phospholipase binds the C-terminal extension of the VgrG trimeric spike. Here, we purify the VgrG-Tle1 complex and show that a VgrG trimer binds three Tle1 monomers and inhibits their activity. Using covalent cross-linking coupled to high-resolution mass spectrometry, we provide information on the sites of contact and further identify the requirement for a Tle1 N-terminal secretion sequence in complex formation. Finally, we report the 2.6-Å-resolution cryo-electron microscopy tri-dimensional structure of the (VgrG) 3-(Tle1) 3 complex revealing how the effector binds its cargo, and how VgrG inhibits Tle1 phospholipase activity. The inhibition of Tle1 phospholipase activity once bound to VgrG suggests that Tle1 dissociation from VgrG is required upon delivery.< Réduire

Mots clés en anglais

type VI secretion system

Structural Biology

Virology & Host Pathogen Interaction

type VI secretion system Subject Categories Microbiology

protein secretion

cryo-electron microscopy

bacterial toxin

bacterial competition

URI

https://oskar-bordeaux.fr/handle/20.500.12278/182445

DOI

http://dx.doi.org/10.15252/embj.2019104129

Projet Européen

Infrastructure for NMR, EM and X-ray crystallography for translational research

Project ANR

Contribution des Effecteurs du Système de Sécrétion de Type VI à la pathogénicité des Escherichia coli adhérents-invasifs - ANR-18-CE15-0013
Guerre bactérienne: Architecture et Fonction du Système de Sécrétion de Type VI - ANR-14-CE14-0006
Bloc-outil et Imagerie de Précision pour le Binage Intra-rang Précoce - ANR-17-ROSE-0001
Cibler le métabolisme des fibroblastes adventitiels activés pour traiter l'hypertension pulmonaire

Unités de recherche

MFP (Laboratoire Microbiologie Fondamentale et Pathogénicité) - UMR 5234

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Structural basis for loading and inhibition of a bacterial T6 SS phospholipase effector by the VgrG spike