Cryo-EM analysis of the SctV cytosolic domain from the enteropathogenic E. coli T3SS injectisome

Abstract

The bacterial injectisome and flagella both rely on type III secretion systems for their assembly. The syringe-like injectisome creates a continuous channel between the bacterium and the host cell, through which signal-modulating effector proteins are secreted. The inner membrane pore protein SctV controls the hierarchy of substrate selection and may also be involved in energizing secretion. We present the 4.7 Å cryo-EM structure of the SctV cytosolic domain (SctV_C) from the enteropathogenic Escherichia coli injectisome. SctV_C forms a nonameric ring with primarily electrostatic interactions between its subunits. Molecular dynamics simulations show that monomeric SctV_C maintains a closed conformation, in contrast with previous studies on flagellar homologue FlhA. Comparison with substrate-bound homologues suggest that a conformational change would be required to accommodate binding partners.

Keywords: Cryo-EM; Export apparatus; Flagella; Injectisome; Molecular dynamics; Type III secretion system.