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

J Antimicrob Chemother. 2014 Jan;69(1):67-71. doi: 10.1093/jac/dkt311. Epub 2013 Aug 8.

Abstract

Objectives: VIM-type enzymes are among the most widespread acquired metallo-β-lactamases among Gram-negative pathogens. Integron In70 is a class 1 integron that has emerged as a successful genetic support for blaVIM-1 (one of the most prevalent blaVIM allelic variants) in Gram-negative non-fermenters, and is 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 pAX22 was 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, In70 was 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 ISPa17 could have in mobilizing defective Tn402-like transposons carrying class 1 integrons.

Keywords: Gram negative; insertion sequence; transposon.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Achromobacter denitrificans / genetics*
  • DNA Transposable Elements
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Gene Order
  • Humans
  • Integrons*
  • Molecular Sequence Data
  • Plasmids*
  • Pseudomonas aeruginosa / genetics
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Synteny
  • beta-Lactamases / genetics

Substances

  • DNA Transposable Elements
  • DNA, Bacterial
  • VIM-1 metallo-beta-lactamase
  • beta-Lactamases

Associated data

  • GENBANK/HF679279