Characterization of plasmid pAX22, encoding VIM-1 metallo-β-lactamase, reveals a new putative mechanism of In70 integron mobilization

Vincenzo Di Pilato, Simona Pollini, Gian Maria Rossolini

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Objectives: VIM-type enzymes are among the most widespread acquired metallo-β-lactamases among Gramnegative pathogens. Integron In70 is a class 1 integron that has emerged as a successful genetic support for blaVIM-1 (one of themostprevalent blaVIM allelic variants) in Gram-negativenon-fermenters, andis usually chromosome borne. The objective of this study was to characterize plasmid pAX22 from Achromobacter xylosoxidans AX22, which represents the only In70-harbouring plasmid known so far, to gather insights into the mechanisms of evolution and dissemination of In70-like elements. Methods: The complete sequence of pAX22was obtained by pyrosequencing and assembled with Roche Newbler software. The draft sequence, completed using a PCR-based strategy, was annotated via the BASys tool and compared with known sequences using BLAST algorithms. Results: The backbone of pAX22 showed significant similarity with that of pNOR-2000, a blaVIM-2-harbouring plasmid from Pseudomonas aeruginosa, and with the TnCP23 transposon. The three elements differed from each other mainly by the class 1 integron cassette arrays and by some integron-associated structures. In pAX22, In70was associated with a novel putative transposon, Tn7017, composed of a defective Tn402-like transposon carrying In70 and the ISPa17 insertion sequence. Conclusion: Plasmid pAX22 belongs to a lineage of plasmids circulating among Gram-negative non-fermenters. In70 was probably acquired by pAX22 by transposition of Tn7017, revealing a novel putative mechanism of In70 mobilization. Our results highlight the potential role that ISPa17couldhave in mobilizing defective Tn402-like transposons carrying class 1 integrons.

Original languageEnglish
Article numberdkt311
Pages (from-to)67-71
Number of pages5
JournalJournal of Antimicrobial Chemotherapy
Volume69
Issue number1
DOIs
Publication statusPublished - Jan 2014

Fingerprint

Integrons
Plasmids
Achromobacter denitrificans
Insertional Mutagenesis
Pseudomonas aeruginosa
Software
Chromosomes
Polymerase Chain Reaction
Enzymes

Keywords

  • Gram negative
  • Insertion sequence
  • Transposon

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Characterization of plasmid pAX22, encoding VIM-1 metallo-β-lactamase, reveals a new putative mechanism of In70 integron mobilization. / Pilato, Vincenzo Di; Pollini, Simona; Rossolini, Gian Maria.

In: Journal of Antimicrobial Chemotherapy, Vol. 69, No. 1, dkt311, 01.2014, p. 67-71.

Research output: Contribution to journalArticle

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abstract = "Objectives: VIM-type enzymes are among the most widespread acquired metallo-β-lactamases among Gramnegative pathogens. Integron In70 is a class 1 integron that has emerged as a successful genetic support for blaVIM-1 (one of themostprevalent blaVIM allelic variants) in Gram-negativenon-fermenters, andis usually chromosome borne. The objective of this study was to characterize plasmid pAX22 from Achromobacter xylosoxidans AX22, which represents the only In70-harbouring plasmid known so far, to gather insights into the mechanisms of evolution and dissemination of In70-like elements. Methods: The complete sequence of pAX22was obtained by pyrosequencing and assembled with Roche Newbler software. The draft sequence, completed using a PCR-based strategy, was annotated via the BASys tool and compared with known sequences using BLAST algorithms. Results: The backbone of pAX22 showed significant similarity with that of pNOR-2000, a blaVIM-2-harbouring plasmid from Pseudomonas aeruginosa, and with the TnCP23 transposon. The three elements differed from each other mainly by the class 1 integron cassette arrays and by some integron-associated structures. In pAX22, In70was associated with a novel putative transposon, Tn7017, composed of a defective Tn402-like transposon carrying In70 and the ISPa17 insertion sequence. Conclusion: Plasmid pAX22 belongs to a lineage of plasmids circulating among Gram-negative non-fermenters. In70 was probably acquired by pAX22 by transposition of Tn7017, revealing a novel putative mechanism of In70 mobilization. Our results highlight the potential role that ISPa17couldhave in mobilizing defective Tn402-like transposons carrying class 1 integrons.",
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N2 - Objectives: VIM-type enzymes are among the most widespread acquired metallo-β-lactamases among Gramnegative pathogens. Integron In70 is a class 1 integron that has emerged as a successful genetic support for blaVIM-1 (one of themostprevalent blaVIM allelic variants) in Gram-negativenon-fermenters, andis usually chromosome borne. The objective of this study was to characterize plasmid pAX22 from Achromobacter xylosoxidans AX22, which represents the only In70-harbouring plasmid known so far, to gather insights into the mechanisms of evolution and dissemination of In70-like elements. Methods: The complete sequence of pAX22was obtained by pyrosequencing and assembled with Roche Newbler software. The draft sequence, completed using a PCR-based strategy, was annotated via the BASys tool and compared with known sequences using BLAST algorithms. Results: The backbone of pAX22 showed significant similarity with that of pNOR-2000, a blaVIM-2-harbouring plasmid from Pseudomonas aeruginosa, and with the TnCP23 transposon. The three elements differed from each other mainly by the class 1 integron cassette arrays and by some integron-associated structures. In pAX22, In70was associated with a novel putative transposon, Tn7017, composed of a defective Tn402-like transposon carrying In70 and the ISPa17 insertion sequence. Conclusion: Plasmid pAX22 belongs to a lineage of plasmids circulating among Gram-negative non-fermenters. In70 was probably acquired by pAX22 by transposition of Tn7017, revealing a novel putative mechanism of In70 mobilization. Our results highlight the potential role that ISPa17couldhave in mobilizing defective Tn402-like transposons carrying class 1 integrons.

AB - Objectives: VIM-type enzymes are among the most widespread acquired metallo-β-lactamases among Gramnegative pathogens. Integron In70 is a class 1 integron that has emerged as a successful genetic support for blaVIM-1 (one of themostprevalent blaVIM allelic variants) in Gram-negativenon-fermenters, andis usually chromosome borne. The objective of this study was to characterize plasmid pAX22 from Achromobacter xylosoxidans AX22, which represents the only In70-harbouring plasmid known so far, to gather insights into the mechanisms of evolution and dissemination of In70-like elements. Methods: The complete sequence of pAX22was obtained by pyrosequencing and assembled with Roche Newbler software. The draft sequence, completed using a PCR-based strategy, was annotated via the BASys tool and compared with known sequences using BLAST algorithms. Results: The backbone of pAX22 showed significant similarity with that of pNOR-2000, a blaVIM-2-harbouring plasmid from Pseudomonas aeruginosa, and with the TnCP23 transposon. The three elements differed from each other mainly by the class 1 integron cassette arrays and by some integron-associated structures. In pAX22, In70was associated with a novel putative transposon, Tn7017, composed of a defective Tn402-like transposon carrying In70 and the ISPa17 insertion sequence. Conclusion: Plasmid pAX22 belongs to a lineage of plasmids circulating among Gram-negative non-fermenters. In70 was probably acquired by pAX22 by transposition of Tn7017, revealing a novel putative mechanism of In70 mobilization. Our results highlight the potential role that ISPa17couldhave in mobilizing defective Tn402-like transposons carrying class 1 integrons.

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