Molecular architecture, polar targeting and biogenesis of the Legionella Dot/Icm T4SS.

Abstract

Legionella pneumophila survives and replicates inside host cells by secreting ~300 effectors through the Dot/Icm type IVB secretion system (T4BSS). Here, we used complementary electron cryotomography (ECT) and immunofluorescence microscopy (IF) to investigate the molecular architecture and biogenesis of the Dot/Icm secretion apparatus. ECT mapped the location of the core and accessory components of the Legionella core-transmembrane subcomplex revealing a well-ordered central channel that opens into a large, windowed secretion chamber with an unusual 13-fold symmetry. IF deciphered an early-stage assembly process that begins with targeting of Dot/Icm components to the bacterial poles. Polar targeting of this T4BSS is mediated by two Dot/Icm proteins, DotU and IcmF, that interestingly are homologs of the T6SS membrane complex components TssL and TssM, suggesting the Dot/Icm T4BSS is a hybrid system. Together these results revealed that the Dot/Icm complex assembles in an “axial-to-peripheral” pattern.