Membrane platform protein PulF of the Klebsiella type II secretion system forms a trimeric ion channel essential for endopilus assembly and protein secretion

• Published in: mBio Vol. 15; no. 1; p. e0142323
• Main Authors: Guilvout, Ingrid, Samsudin, Firdaus, Huber, Roland G, Bond, Peter J, Bardiaux, Benjamin, Francetic, Olivera
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 16.01.2024
• Subjects: cardiolipin; ion channel; Life Sciences; Microbiology and Parasitology; Molecular and Cellular Biology; Research Article; residue coevolution; type II secretion system; type IV filaments; type IV pili; type II secretion system; ion channel; AlphaFold2; residue coevolution; type IV filaments; cardiolipin; type IV pili; ion motive force
• ISSN: 2150-7511; 2161-2129; 2150-7511
• DOI: 10.1128/mbio.01423-23

Type IV pili and type II secretion systems are members of the widespread type IV filament (T4F) superfamily of nanomachines that assemble dynamic and versatile surface fibers in archaea and bacteria. The assembly and retraction of T4 filaments with diverse surface properties and functions require the plasma membrane platform proteins of the GspF/PilC superfamily. Generally considered dimeric, platform proteins are thought to function as passive transmitters of the mechanical energy generated by the ATPase motor, to somehow promote insertion of pilin subunits into the nascent pilus fibers. Here, we generate and experimentally validate structural predictions that support the trimeric state of a platform protein PulF from a type II secretion system. The PulF trimers form selective proton or sodium channels which might energize pilus assembly using the membrane potential. The conservation of the channel sequence and structural features implies a common mechanism for all T4F assembly systems. We propose a model of the oligomeric PulF-PulE ATPase complex that provides an essential framework to investigate and understand the pilus assembly mechanism.