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Journal of Bacteriology, February 2009, p. 1239-1247, Vol. 191, No. 4
0021-9193/09/$08.00+0     doi:10.1128/JB.01071-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Conjugative Interaction Induces Transposition of ISPst9 in Pseudomonas stutzeri AN10{triangledown}

J. A. Christie-Oleza,1 M. P. Lanfranconi,1 B. Nogales,1 J. Lalucat,1,2 and R. Bosch1*

Microbiologia, Departament de Biologia, Universitat de les Illes Balears (UIB), Palma de Mallorca, Spain,1 Institut Mediterrani d'Estudis Avançats (IMEDEA), Universitat de les Illes Balears (UIB), Palma de Mallorca, Spain2

Received 31 July 2008/ Accepted 30 November 2008

ISPst9 is an ISL3-like insertion sequence (IS) that was recently described in the naphthalene-degrading organism Pseudomonas stutzeri strain AN10. In this paper we describe a novel strong IS regulation stimulus; transposition of ISPst9 is induced in all P. stutzeri AN10 cells after conjugative interaction with Escherichia coli. Thus, we observed that in all P. stutzeri AN10 cells that received genetic material by conjugation the ISPst9 genomic dose and/or distribution was changed. Furthermore, ISPst9 transposition was also observed when P. stutzeri AN10 cells were put in contact with the plasmidless conjugative strain E. coli S17-1{lambda}pir, but not when they were put in contact with E. coli DH5{alpha} (a nonconjugative strain). The mechanism of ISPst9 transposition was analyzed, and transposition was shown to proceed by excision from the donor DNA using a conservative mechanism, which generated 3- to 10-bp deletions of the flanking DNA. Our results indicate that ISPst9 transposes, forming double-stranded DNA circular intermediates consisting of the IS and a 5-bp intervening DNA sequence probably derived from the ISPst9 flanking regions. The kinetics of IS circle formation are also described.

* Corresponding author. Mailing address: Microbiologia, Departament de Biología, Universitat de les Illes Balears, Carretera Valldemossa, km 7.5, 07122 Palma de Mallorca, Spain. Phone: 34-971-172738. Fax: 34-971-173184. E-mail: rbosch{at}uib.es

{triangledown} Published ahead of print on 5 December 2008.

Journal of Bacteriology, February 2009, p. 1239-1247, Vol. 191, No. 4
0021-9193/09/$08.00+0     doi:10.1128/JB.01071-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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