title: Calcium binding properties of the Kingella kingae PilC1 and PilC2 proteins have differential effects on type IV pilus-mediated adherence and twitching motility.
abstract: Kingella kingae is an emerging bacterial pathogen that is being recognized increasingly as an important etiology of septic arthritis, osteomyelitis, and bacteremia, especially in young children. The pathogenesis of K. kingae disease begins with bacterial adherence to respiratory epithelium, which is dependent on type IV pili and is influenced by two PilC-like proteins called PilC1 and PilC2. Production of either PilC1 or PilC2 is necessary for K. kingae piliation and bacterial adherence. In this study, we set out to further investigate the role of PilC1 and PilC2 in type IV pilus-associated phenotypes. We found that PilC1 contains a functional 9-amino-acid calcium-binding (Ca-binding) site with homology to the Pseudomonas aeruginosa PilY1 Ca-binding site and that PilC2 contains a functional 12-amino-acid Ca-binding site with homology to the human calmodulin Ca-binding site. Using targeted mutagenesis to disrupt the Ca-binding sites, we demonstrated that the PilC1 and PilC2 Ca-binding sites are dispensable for piliation. Interestingly, we showed that the PilC1 site is necessary for twitching motility and adherence to Chang epithelial cells, while the PilC2 site has only a minor influence on twitching motility and no influence on adherence. These findings establish key differences in PilC1 and PilC2 function in K. kingae and provide insights into the biology of the PilC-like family of proteins.
authors.affiliation: Department of Pediatrics, Duke University Medical Center, Durham, NC, USA.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Porsch
authors.forename: Michael D L
authors.initials: MD
authors.lastname: Johnson
authors.forename: Angela D
authors.initials: AD
authors.lastname: Broadnax
authors.forename: Christopher K
authors.initials: CK
authors.lastname: Garrett
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Redinbo
authors.forename: Joseph W
authors.initials: JW
authors.lastname: St Geme
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology*; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000145:classification / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D017882:Kingella kingae / Q000235:genetics / Q000378:metabolism*; D009068:Movement; D010641:Phenotype; D010957:Plasmids; D011485:Protein Binding
chemical_list: D036781:Fimbriae Proteins; D002118:Calcium