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Infection and Immunity, February 1999, p. 717-725, Vol. 67, No. 2
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The R-Type Pyocin of Pseudomonas aeruginosa C Is a Bacteriophage Tail-Like Particle That Contains Single-Stranded DNA

Frank K. N. Lee,1 Kathleen C. Dudas,2 Julie A. Hanson,3 M. Bud Nelson,4 Philip T. LoVerde,5 and Michael A. Apicella1,*

Department of Microbiology, College of Medicine, University of Iowa, Iowa City, Iowa1; Department of Microbiology, Duke University, Durham, North Carolina2; Department of Internal Medicine---Nephrology, University of Michigan Hospital, Ann Arbor, Michigan3; BioCrystal, Westerville, Ohio4; and Department of Microbiology, State University of New York at Buffalo, Buffalo, New York5

Received 23 July 1998/Returned for modification 6 October 1998/Accepted 17 November 1998

Pseudomonas aeruginosa R-type pyocin particles have been described as bacteriocins that resemble bacteriophage tail-like structures. Because of their unusual structure, we reexamined whether they contained nucleic acids. Our data indicated that pyocin particles isolated from P. aeruginosa C (pyocin C) contain DNA. Probes generated from this DNA by the random-primer extension method hybridized to distinct bands in restriction endonuclease-digested P. aeruginosa C genomic DNA. These probes also hybridized to genomic DNA from 6 of 18 P. aeruginosa strains that produced R-type pyocins. Asymmetric PCR, complementary oligonucleotide hybridization, and electron microscopy indicated that pyocin C particles contained closed circular single-stranded DNA, approximately 4.0 kb in length. Examination of total intracellular DNA from mitomycin C-induced cultures revealed the presence of two extrachromosomal DNA molecules, a double-stranded molecule and a single-stranded molecule, which hybridized to pyocin DNA. Sequence analysis of 7,480 nucleotides of P. aeruginosa C chromosomal DNA containing the pyocin DNA indicated the presence of pyocin open reading frames with similarities to open reading frames from filamentous phages and cryptic phage elements. We did not observe any similarities to known phage structural proteins or previously characterized pseudomonal prt genes expressing R-type pyocin structural proteins. These studies demonstrate that pyocin particles from P. aeruginosa C are defective phages that contain a novel closed circular single-stranded DNA and that this DNA was derived from the chromosome of P. aeruginosa C.

* Corresponding author. Mailing address: 3-403 BSB, Department of Microbiology, College of Medicine, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7807. Fax: (319) 335-9006. E-mail: michael-apicella{at}uiowa.edu.

Infection and Immunity, February 1999, p. 717-725, Vol. 67, No. 2
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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