title: The peptidoglycan-binding protein FimV promotes assembly of the Pseudomonas aeruginosa type IV pilus secretin.
abstract: The Pseudomonas aeruginosa inner membrane protein FimV is among several proteins of unknown function required for type IV pilus-mediated twitching motility, arising from extension and retraction of pili from their site of assembly in the inner membrane. The pili transit the periplasm and peptidoglycan (PG) layer, ultimately exiting the cell through the PilQ secretin. Although fimV mutants are nonmotile, they are susceptible to killing by pilus-specific bacteriophage, a hallmark of retractable surface pili. Here we show that levels of recoverable surface pili were markedly decreased in fimV pilT retraction-deficient mutants compared with levels in the pilT control, demonstrating that FimV acts at the level of pilus assembly. Levels of inner membrane assembly subcomplex proteins PilM/N/O/P were decreased in fimV mutants, but supplementation of these components in trans did not restore pilus assembly or motility. Loss of FimV dramatically reduced the levels of the PilQ secretin multimer through which pili exit the cell, in part due to decreased levels of PilQ monomers, while PilF pilotin levels were unchanged. Expression of pilQ in trans in the wild type or fimV mutants increased total PilQ monomer levels but did not alter secretin multimer levels or motility. PG pulldown assays showed that the N terminus of FimV bound PG in a LysM motif-dependent manner, and a mutant with an in-frame chromosomal deletion of the LysM motif had reduced motility, secretin levels, and surface piliation. Together, our data show that FimV's role in pilus assembly is to promote secretin formation and that this function depends upon its PG-binding domain.
authors.affiliation: Department of Biochemistry and Biomedical Sciences, Michael G. DeGroote Institute for Infectious Diseases Research, University McMaster University, Hamilton, ON, Canada.
authors.forename: Hania
authors.initials: H
authors.lastname: Wehbi
authors.forename: Eder
authors.initials: E
authors.lastname: Portillo
authors.forename: Hanjeong
authors.initials: H
authors.lastname: Harvey
authors.forename: Anthony E
authors.initials: AE
authors.lastname: Shimkoff
authors.forename: Edie M
authors.initials: EM
authors.lastname: Scheurwater
authors.forename: P Lynne
authors.initials: PL
authors.lastname: Howell
authors.forename: Lori L
authors.initials: LL
authors.lastname: Burrows
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D008124:Locomotion; D010457:Peptidoglycan / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D011550:Pseudomonas aeruginosa / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan; C084295:pilQ protein, bacteria; D036781:Fimbriae Proteins