title: Functional mapping of PilF and PilQ in the Pseudomonas aeruginosa type IV pilus system.
abstract: Pseudomonas aeruginosa uses type IV pili (T4P) to interact with the environment and as key virulence factors when acting as an opportunistic pathogen. Assembly of the outer membrane PilQ secretin channel through which the pili are extruded is essential for pilus biogenesis. The P. aeruginosa T4P pilotin, PilF, is required for PilQ outer membrane localization and assembly into secretins and contains six tetratricopeptide (TPR) protein-protein interaction motifs, suggesting that the two proteins interact. In this study, we found that the first four TPR motifs of PilF are sufficient for PilQ outer membrane targeting, oligomerization, and function. Guided by our structure of PilF, site-directed mutagenesis of the protein surface revealed that a hydrophobic groove on the first TPR is required for PilF-mediated PilQ assembly. Deletion of individual domains within PilQ suggests that the N0, KH-like, or secretin domain, but not the C-terminus, interacts with PilF. Purified PilQ was found to pull down PilF from Pseudomonas cell lysates. Together, these data allow us to propose a model for PilF function in the T4P system. PilF interacts directly or indirectly with the PilQ monomer after translocation of both proteins through the inner membrane and acts as a co-chaperone with the Lol system to facilitate transit across the periplasm to the outer membrane. The mechanism of PilQ insertion and assembly, which appears to be independent of the Bam system, remains to be determined.
authors.affiliation: Program in Molecular Structure and Function, Research Institute, Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: Jason
authors.initials: J
authors.lastname: Koo
authors.forename: Tim
authors.initials: T
authors.lastname: Tang
authors.forename: Hanjeong
authors.initials: H
authors.lastname: Harvey
authors.forename: Stephanie
authors.initials: S
authors.lastname: Tammam
authors.forename: Liliana
authors.initials: L
authors.lastname: Sampaleanu
authors.forename: Lori L
authors.initials: LL
authors.lastname: Burrows
authors.forename: P Lynne
authors.initials: PL
authors.lastname: Howell
mesh_terms: D036781:Fimbriae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial; D005816:Genetic Complementation Test; D008958:Models, Molecular; D016296:Mutagenesis; D011550:Pseudomonas aeruginosa / Q000378:metabolism*
chemical_list: C102919:PilF protein, Pseudomonas; C084295:pilQ protein, bacteria; D036781:Fimbriae Proteins