Pseudomonas aeruginosa Type IV pilus expression in Neisseria gonorrhoeae: effects of pilin subunit composition on function and organelle dynamics.
Type IV pili (TFP) play central roles in the expression of many phenotypes including motility, multicellular behavior, sensitivity to bacteriophages, natural genetic transformation, and adherence. In Neisseria gonorrhoeae, these properties require ancillary proteins that act in conjunction with TFP expression and influence organelle dynamics. Here, the intrinsic contributions of the pilin protein itself to TFP dynamics and associated phenotypes were examined by expressing the Pseudomonas aeruginosa PilA(PAK) pilin subunit in N. gonorrhoeae. We show here that, although PilA(PAK) pilin can be readily assembled into TFP in this background, steady-state levels of purifiable fibers are dramatically reduced relative those of endogenous pili. This defect is due to aberrant TFP dynamics as it is suppressed in the absence of the PilT pilus retraction ATPase. Functionally, PilA(PAK) pilin complements gonococcal adherence for human epithelial cells but only in a pilT background, and this property remains dependent on the coexpression of both the PilC adhesin and the PilV pilin-like protein. Since P. aeruginosa pilin only moderately supports neisserial sequence-specific transformation despite its assembly proficiency, these results together suggest that PilA(PAK) pilin functions suboptimally in this environment. This appears to be due to diminished compatibility with resident proteins essential for TFP function and dynamics. Despite this, PilA(PAK) pili support retractile force generation in this background equivalent to that reported for endogenous pili. Furthermore, PilA(PAK) pili are both necessary and sufficient for bacteriophage PO4 binding, although the strain remains phage resistant. Together, these findings have significant implications for TFP biology in both N. gonorrhoeae and P. aeruginosa.
Winther-Larsen
Hanne C
HC
Department of Molecular Biosciences, University of Oslo, P.O. Box 1041 Blindern, 0317 Oslo, Norway. hannewi@biotek.uio.no
Wolfgang
Matthew C
MC
van Putten
Jos P M
JP
Roos
Norbert
N
Aas
Finn Erik
FE
Egge-Jacobsen
Wolfgang M
WM
Maier
Berenike
B
Koomey
Michael
M
D001422:Bacterial Adhesion; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology / Q000382:microbiology; D004847:Epithelial Cells / Q000382:microbiology; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D046529:Microscopy, Electron, Transmission; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000502:physiology; D010641:Phenotype; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*
D036781:Fimbriae Proteins