Direct interactions between the secreted effector and the T2SS components GspL and GspM reveal a new effector-sensing step during type 2 secretion

Sandra Michel-Souzy (1) , Badreddine Douzi (1, 2) , Frederic Cadoret (1) , Claire Raynaud (1, 2) , Loïc Quinton (3) , Genevieve Ball (1, 2) , Romé Voulhoux (2, 1)

1 LISM - Laboratoire d'ingénierie des systèmes macromoléculaires
2 LCB - Laboratoire de chimie bactérienne
3 Laboratoire de Spectrométrie de Masse

In many Gram-negative bacteria, the type 2 secretion system (T2SS) plays an important role in virulence because of its capacity to deliver a large amount of fully folded protein effectors to the extracellular milieu. Despite our knowledge of most T2SS components, the mechanisms underlying effector recruitment and secretion by the T2SS remain enigmatic. Using complementary biophysical and biochemical approaches, we identified here two direct interactions between the secreted effector CbpD and two components, XcpY L and XcpZ M , of the T2SS assembly platform (AP) in the opportunistic pathogen Pseudomonas aeruginosa. Competition experiments indicated that CbpD binding to XcpY L is XcpZ M-dependent, suggesting sequential recruitment of the effector by the periplasmic domains of these AP components. Using the bacterial two-hybrid system, we then tested the influence of the effector on the AP protein-protein interaction network. Our findings revealed that the presence of the effector modifies the AP interactome and, in particular, induces XcpZ M homodimerization and increases the affinity between XcpY L and XcpZ M. The observed direct relationship between effector binding and T2SS dynamics suggests an additional synchronizing step during the type 2 secretion process, where the activation of the AP of the T2SS nanomachine is triggered by effector binding.

Mots clés: Xcp; effector recognition; Pseudomonas aeruginosa; CbpD; virulence; infectious disease; Protein Secretion; Type 2 Secretion; T2SS; Gsp

Domaines: Sciences du Vivant [q-bio]

Sandra Michel-Souzy, Badreddine Douzi, Frederic Cadoret, Claire Raynaud, Loïc Quinton, et al.. Direct interactions between the secreted effector and the T2SS components GspL and GspM reveal a new effector-sensing step during type 2 secretion. Journal of Biological Chemistry, 2018, 293 (50), pp.19441-19450. ⟨10.1074/jbc.RA117.001127⟩. ⟨hal-01919531⟩