Coevolution-guided mapping of the Type VI secretion membrane complexbaseplate interface

Etienne Vanlioğlu (1) , Yoann Santin (1) , Isaac Filella-Merce (2) , Riccardo Pellarin (2) , Eric Cascales (1)

1 LISM - Laboratoire d'ingénierie des systèmes macromoléculaires
2 Bioinformatique structurale - Structural Bioinformatics

The type VI secretion system (T6SS) is a multiprotein weapon evolved by Gram-negative bacteria to deliver effectors into eukaryotic cells or bacterial rivals. The T6SS uses a contractile mechanism to propel an effector-loaded needle into its target. The contractile tail is built on an assembly platform, the baseplate, which is anchored to a membrane complex. Baseplate-membrane complex interactions are mainly mediated by contacts between the C-terminal domain of the TssK baseplate component and the cytoplasmic domain of the TssL inner membrane protein. Currently, the structural details of this interaction are unknown due to the marginal stability of the TssK-TssL complex. Here we conducted a mutagenesis study based on putative TssK-TssL contact pairs identified by co-evolution analyses. We then evaluated the impact of these mutations on T6SS activity, TssK-TssL interaction and sheath assembly and dynamics in enteroaggregative Escherichia coli. Finally, we probed the TssK-TssL interface by disulfide cross-linking, allowing to propose a model for the baseplate-membrane complex interface.

Mots clés: protein transport; bacterial competition; protein-protein interaction; coevolution; contact sites; protein transport

Domaines: Sciences du Vivant [q-bio]

Etienne Vanlioğlu, Yoann Santin, Isaac Filella-Merce, Riccardo Pellarin, Eric Cascales. Coevolution-guided mapping of the Type VI secretion membrane complexbaseplate interface. Journal of Molecular Biology, 2023, 435 (2), pp.167918. ⟨10.1016/j.jmb.2022.167918⟩. ⟨hal-04234783⟩