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Antimicrobial Agents and Chemotherapy, April 2008, p. 1472-1480, Vol. 52, No. 4
0066-4804/08/$08.00+0     doi:10.1128/AAC.00982-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Different Pathways to Acquiring Resistance Genes Illustrated by the Recent Evolution of IncW Plasmids{triangledown} ,{dagger}

Carlos Revilla,1,# M. Pilar Garcillán-Barcia,1,# Raúl Fernández-López,1 Nicholas R. Thomson,2 Mandy Sanders,2 Martin Cheung,3 Christopher M. Thomas,3 and Fernando de la Cruz1*

Departamento de Biología Molecular e Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC-IDICAN, C. Herrera Oria s/n, 39011 Santander, Spain,1 The Pathogen Sequencing Unit, Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge CB10 1SA, United Kingdom,2 School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom3

Received 28 July 2007/ Returned for modification 23 September 2007/ Accepted 3 February 2008

DNA sequence analysis of five IncW plasmids (R388, pSa, R7K, pIE321, and pIE522) demonstrated that they share a considerable portion of their genomes and allowed us to define the IncW backbone. Among these plasmids, the backbone is stable and seems to have diverged recently, since the overall identity among its members is higher than 95%. The only gene in which significant variation was observed was trwA; the changes in the coding sequence correlated with parallel changes in the corresponding TrwA binding sites at oriT, suggesting a functional connection between both sets of changes. The present IncW plasmid diversity is shaped by the acquisition of antibiotic resistance genes as a consequence of the pressure exerted by antibiotic usage. Sequence comparisons pinpointed the insertion events that differentiated the five plasmids analyzed. Of greatest interest is that a single acquisition of a class I integron platform, into which different gene cassettes were later incorporated, gave rise to plasmids R388, pIE522, and pSa, while plasmids R7K and pIE321 do not contain the integron platform and arose in the antibiotic world because of the insertion of several antibiotic resistance transposons.

* Corresponding author. Mailing address: Departamento de Biología Molecular e Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC-IDICAN, C. Herrera Oria s/n, 39011 Santander, Spain. Phone: 34-942-201942. Fax: 34-942-201945. E-mail: delacruz{at}unican.es

{triangledown} Published ahead of print on 11 February 2008.

{dagger} Supplemental material for this article may be found at http://aac.asm.org/.

# These authors are equal contributors.

Antimicrobial Agents and Chemotherapy, April 2008, p. 1472-1480, Vol. 52, No. 4
0066-4804/08/$08.00+0     doi:10.1128/AAC.00982-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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