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Journal of Bacteriology, March 2005, p. 1740-1750, Vol. 187, No. 5
0021-9193/05/$08.00+0     doi:10.1128/JB.187.5.1740-1750.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Comparative Study of Class 1 Integron and Vibrio cholerae Superintegron Integrase Activities

Latefa Biskri,1,{dagger} Marie Bouvier,1 Anne-Marie Guérout,1 Stéphanie Boisnard,1,{ddagger} and Didier Mazel1*

Unité Postulante Plasticité du Génome Bactérien, CNRS URA 2171, Département Structure et Dynamique des Génomes, Institut Pasteur, Paris, France1

Received 15 July 2004/ Accepted 18 November 2004

Superintegrons (SIs) and multiresistant integrons (MRIs) have two main structural differences: (i) the SI platform is sedentary, while the MRI platform is commonly associated with mobile DNA elements and (ii) the recombination sites (attC) of SI gene cassette clusters are highly homogeneous, while those of MRI cassette arrays are highly variable in length and sequence. In order to determine if the latter difference was correlated with a dissimilarity in the recombination activities, we conducted a comparative study of the integron integrases of the class 1 MRI (IntI1) and the Vibrio cholerae SI (VchIntIA). We developed two assays that allowed us to independently measure the frequencies of cassette deletion and integration at the cognate attI sites. We demonstrated that the range of attC sites efficiently recombined by VchIntIA is narrower than the range of attC sites efficiently recombined by IntI1. Introduction of mutations into the V. cholerae repeats (VCRs), the attC sites of the V. cholerae SI cassettes, allowed us to map positions that affected the VchIntIA and IntI1 activities to different extents. Using a cointegration assay, we established that in E. coli, attI1-x-VCR recombination catalyzed by IntI1 was 2,600-fold more efficient than attIVch-x-VCR recombination catalyzed by VchIntIA. We performed the same experiments in V. cholerae and established that the attIVch-x-VCR recombination catalyzed by VchIntIA was 2,000-fold greater than the recombination measured in E. coli. Taken together, our results indicate that in the V. cholerae SI, the substrate recognition and recombination reactions mediated by VchIntIA might differ from the class 1 MRI paradigm.

* Corresponding author. Mailing address: Unité Postulante Plasticité du Génome Bactérien, CNRS URA 2171, Institut Pasteur, 25 rue du Dr Roux, 75724, Paris, France. Phone: 33 1 40 61 32 84. Fax: 33 1 45 68 87 90. E-mail: mazel{at}pasteur.fr.

{dagger} Present address: Laboratoire de Bactériologie Moléculaire, Faculté de Médecine, Université Libre de Bruxelles, B-1070 Brussels, Belgium.

{ddagger} Present address: Institut de Génétique et Microbiologie, Université Paris-Sud, UMR 8621, 91405 Orsay Cedex, France.

Journal of Bacteriology, March 2005, p. 1740-1750, Vol. 187, No. 5
0021-9193/05/$08.00+0     doi:10.1128/JB.187.5.1740-1750.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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