Conjugative mobilization of a vancomycin resistance plasmid by a putative Enterococcus faecium sex pheromone response plasmid

Recent epidemiological evidence suggests that horizontal gene transfer may be an important mechanism for dissemination of vancomycin resistance, A filter mating survey of 21 VanA Enterococcus faecium isolates from The New York Hospital showed that 14 of these isolates transferred vancomycin resistance (Vm(r)) to the plasmid-free reference strain Enterococcus faecalis JH2-2. One isolate, E. faecium R7, was selected for further study based on its ability to transfer Vm(r) to strain JH2-2 in liquid culture, Analysis of the plasmid content of transconjugants revealed three general classes, The predominant class (28 of 47 transconjugants) contained two separate plasmids: pHKK702 and pHKK703. pHKK702 is a 41-kb plasmid that contains an element indistinguishable from the Vm(r) transposon Tn1546 and an element that hybridizes with an ermB probe from the Staphylococcus aureus erythromycin resistance transposon Tn551. pHKK703 is a 55-kb plasmid that hybridizes with probes for the sex pheromone response genes prgA, prgB, and prgX derived from the E. faecalis plasmid pCF10, The second group of transconjugants (18 of 47) contained various recombinant forms of pHKK702 and pHKK703, whereas a third transconjugant class contained only pHKK702 (1 of 47), Transconjugants that contained both pHKK702 and pHKK703 were able to efficiently transfer Vm(r) to recipient strains in broth or on filters, However, no transfer of Vm(r) was detected using the donor containing only pHKK702. The transfer of Vm(r) from the recombination-deficient derivative of E. faecalis JH2-2 [strain UV202(pHKK702, pHKK703)] was reduced 600-fold compared to that of JH2-2(pHKK702, pHKK703). We propose that pHKK703 functions as an E. faecium sex pheromone response plasmid that conjugatively mobilizes pHKK702, and that a major pathway for this mobilization may require donor-mediated recombination proficiency, This report provides the first example in which a plasmid containing a Tn1546-related element is conjugatively mobilized.