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Journal of Bacteriology, April 2000, p. 2170-2178, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Replication of Staphylococcal Multiresistance Plasmids

Neville Firth,1 Sumalee Apisiridej,1,dagger Tracey Berg,1 Brendon A. O'Rourke,1 Steve Curnock,2 Keith G. H. Dyke,2 and Ronald A. Skurray1,*

School of Biological Sciences, University of Sydney, Sydney, New South Wales 2006, Australia,1 and Microbiology Unit, Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom2

Received 24 November 1999/Accepted 21 January 2000

Based on structural and functional properties, three groups of large staphylococcal multiresistance plasmids have been recognized, viz., the pSK1 family, pSK41-like conjugative plasmids, and beta -lactamase-heavy-metal resistance plasmids. Here we describe an analysis of the replication functions of a representative of each of these plasmid groups. The replication initiation genes from the Staphylococcus aureus plasmids pSK1, pSK41, and pI9789::Tn552 were found to be related to each other and to the Staphylococcus xylosus plasmid pSX267 and are also related to rep genes of several plasmids from other gram-positive genera. Nucleotide sequence similarity between pSK1 and pI9789::Tn552 extended beyond their rep genes, encompassing upstream divergently transcribed genes, orf245 and orf256, respectively. Our analyses revealed that genes encoding proteins related to the deduced orf245 product are variously represented, in several types of organization, on plasmids possessing six seemingly evolutionarily distinct types of replication initiation genes and including both theta-mode and rolling-circle replicons. Construction of minireplicons and subsequent functional analysis demonstrated that orf245 is required for the segregational stability of the pSK1 replicon. In contrast, no gene equivalent to orf245 is evident on the conjugative plasmid pSK41, and a minireplicon encoding only the pSK41 rep gene was found to exhibit a segregational stability approaching that of the parent plasmid. Significantly, the results described establish that many of the large multiresistance plasmids that have been identified in clinical staphylococci, which were formerly presumed to be unrelated, actually utilize an evolutionarily related theta-mode replication system.

* Corresponding author. Mailing address: School of Biological Sciences, University of Sydney, Sydney, New South Wales 2006, Australia. Phone: 61 2 9351-2376. Fax: 61 2 9351-4771. E-mail: skurray{at}bio.usyd.edu.au.

dagger Present address: Division of Medical Microbiology, Department of Pathology, Faculty of Medicine, Prince of Songkla University, Hadyai 90112, Thailand.

Journal of Bacteriology, April 2000, p. 2170-2178, Vol. 182, No. 8
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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