Abstract

Transposons of the Pseudomonas aeruginosa accessory gene pool contribute to phenotype and to genome plasticity. We studied local P. aeruginosa strains to ascertain the encroachment of mer-type res site hunter transposons into clinical settings and their associations with other functional modules. Five different Tn5053 family transposons were detected, all chromosomal. Some were solitary elements; one was in res of Tn1013#, a relative of a reported carrier of int-type res site hunters (class 1 integrons), but most were in res of Tn6603, a new Tn501-related transposon of unknown phenotype. Most of the Tn6603::Tn elements, and some Tn6603 and Tn6603::Tn elements found in GenBank sequences, were at identical sites in an hypothetical gene of P. aeruginosa genomic island PAGI-5v. The island in clonally differing strains was at either of two tRNALys loci, suggesting lateral transfer to these sites. This observation is consistent with the membership of the prototype PAGI-5 island to the ICE family of mobile genetic elements. Additionally, the res site hunters in the nested transposons occupied different positions in the Tn6603 carrier. This suggested independent insertion events on five occasions at least. Tn5053 family members that were mer-/tni-defective were found in Tn6603- and Tn501-like carriers in GenBank sequences of non-clinical Pseudomonas spp. The transposition events in these cases presumably utilized tni functions in trans, as can occur with class 1 integrons. We suggest that in the clinical context, P. aeruginosa strains that carry Tn6603 alone or in PAGI-5v can serve to disseminate functional res site hunters; these in turn can provide the requisite trans-acting tni functions to assist in the dissemination of class 1 integrons, and hence of their associated antibiotic resistance determinants.

Highlights

  • Pseudomonas aeruginosa is a metabolically versatile bacterium that inhabits many diverse ecological niches and is a significant opportunistic nosocomial pathogen in humans [1,2]

  • The inherited genes are frequently located within mobile genetic elements (MGEs), combinations of which contribute to the accessory gene pool of individual strains and aid in niche adaptation [3]

  • For selection of E. coli strains, antimicrobials were added to Nutrient agar (NA) at the following final concentrations: carbenicillin (Cb) at 100 or 250; chloramphenicol (Cm) at 10; mercuric chloride (Hg) at 15; nalidixic acid (Nal) at 8; streptomycin (Sm) at 100; tetracycline hydrochloride (Tc) at 10 or 80

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Summary

Introduction

Pseudomonas aeruginosa is a metabolically versatile bacterium that inhabits many diverse ecological niches and is a significant opportunistic nosocomial pathogen in humans [1,2]. The rich repertoire of MGEs in P. aeruginosa includes plasmids, transposons (Tn), genomic islands (GIs), integrative and conjugative elements (ICEs), and integrons (In), which are often present in strains that exhibit combined resistance to mercury(II) (mer) and to antibiotics. The former phenotype is associated with mer transposons [4,5] or independent mer modules, for example in ICEs and GIs [6,7,8], or in combination with antibiotic-resistance genes, such as occurs in the prevalent IncP-2 plasmids and the ubiquitous IncP-1 plasmids that occur widely in Gram-negative bacteria [9,10,11,12]. This group has the capacity to aid in the spread of class 1 integrons [15,16], which are prominent members of the Tn402 family and encode a multiplicity of antibiotic-resistant gene cassettes [17,18]

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