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Pseudomonas aeruginosa Type IV Pilus Expression in Neisseria gonorrhoeae: Effects of Pilin Subunit Composition on Function and Organelle Dynamics ^,

Hanne C. Winther-Larsen,^1,2* Matthew C. Wolfgang,³ Jos P. M. van Putten,⁴ Norbert Roos,² Finn Erik Aas,^1,2 Wolfgang M. Egge-Jacobsen,^1,2 Berenike Maier,^5, and Michael Koomey^1,2

Centre for Molecular Biology and Neuroscience,¹ Department of Molecular Biosciences, University of Oslo, 0316 Oslo, Norway,² Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599,³ Department of Infectious Diseases and Immunology, Utrecht University, NL-3584 CL, Utrecht, The Netherlands,⁴ Department of Biological Sciences, Columbia University, New York, New York 10027⁵

Received 20 March 2007/ Accepted 6 June 2007

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.

Published ahead of print on 15 June 2007.

Supplemental material for this article may be found at http://jb.asm.org/.

Present address: Westfälische Wilhelms-Universität Münster, Institut für Allgemeine Zoologie und Genetik, 48149 Münster, Germany.