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Journal of Bacteriology, February 2005, p. 1455-1464, Vol. 187, No. 4
0021-9193/05/$08.00+0     doi:10.1128/JB.187.4.1455-1464.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

DNA Binding: a Novel Function of Pseudomonas aeruginosa Type IV Pili

Erin J. van Schaik,1 Carmen L. Giltner,1 Gerald F. Audette,1 David W. Keizer,2,{dagger} Daisy L. Bautista,1 Carolyn M. Slupsky,2,{ddagger} Brian D. Sykes,2 and Randall T. Irvin1*

Department of Medical Microbiology and Immunology,1 Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada2

Received 7 August 2004/ Accepted 14 October 2004

The opportunistic pathogen Pseudomonas aeruginosa produces multifunctional, polar, filamentous appendages termed type IV pili. Type IV pili are involved in colonization during infection, twitching motility, biofilm formation, bacteriophage infection, and natural transformation. Electrostatic surface analysis of modeled pilus fibers generated from P. aeruginosa strain PAK, K122-4, and KB-7 pilin monomers suggested that a solvent-exposed band of positive charge may be a common feature of all type IV pili. Several functions of type IV pili, including natural transformation and biofilm formation, involve DNA. We investigated the ability of P. aeruginosa type IV pili to bind DNA. Purified PAK, K122-4, and KB-7 pili were observed to bind both bacterial plasmid and salmon sperm DNA in a concentration-dependent and saturable manner. PAK pili had the highest affinity for DNA, followed by K122-4 and KB-7 pili. DNA binding involved backbone interactions and preferential binding to pyrimidine residues even though there was no evidence of sequence-specific binding. Pilus-mediated DNA binding was a function of the intact pilus and thus required elements present in the quaternary structure. However, binding also involved the pilus tip as tip-specific, but not base-specific, antibodies inhibited DNA binding. The conservation of a Thr residue in all type IV pilin monomers examined to date, along with the electrostatic data, implies that DNA binding is a conserved function of type IV pili. Pilus-mediated DNA binding could be important for biofilm formation both in vivo during an infection and ex vivo on abiotic surfaces.

* Corresponding author. Mailing address: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada. Phone: (780) 492-5374. Fax: (780) 492-7521. E-mail: Randy.Irvin{at}ualberta.ca.

{dagger} Present address: The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.

{ddagger} Present address: Chenomx Inc., Edmonton, Alberta, Canada.

Journal of Bacteriology, February 2005, p. 1455-1464, Vol. 187, No. 4
0021-9193/05/$08.00+0     doi:10.1128/JB.187.4.1455-1464.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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