Evolution of a 72-Kilobase Cointegrant, Conjugative Multiresistance Plasmid in Community-Associated Methicillin-Resistant Staphylococcus aureus Isolates from the Early 1990s.
Karina Yui Eto, Neville Firth, Amy M Davis, Stephen M Kwong, Marcelina Krysiak, Yung Thin Lee, Frances G O'Brien, Warren B Grubb, Geoffrey W Coombs, Charles S Bond, Joshua P Ramsay
Author Information
Karina Yui Eto: School of Molecular Sciences, University of Western Australia, Crawley, WA, Australia.
Neville Firth: School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia.
Amy M Davis: Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
Stephen M Kwong: School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia.
Marcelina Krysiak: Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
Yung Thin Lee: Antimicrobial Resistance and Infectious Disease Research Laboratory, Murdoch University, Perth, WA, Australia.
Frances G O'Brien: Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
Warren B Grubb: Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
Geoffrey W Coombs: Antimicrobial Resistance and Infectious Disease Research Laboratory, Murdoch University, Perth, WA, Australia.
Charles S Bond: School of Molecular Sciences, University of Western Australia, Crawley, WA, Australia.
Joshua P Ramsay: Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia joshramsay@gmail.com. ORCID
Horizontal transfer of plasmids encoding antimicrobial resistance and virulence determinants has been instrumental in evolution, including the emergence of community-associated methicillin-resistant (CA-MRSA). In the early 1990s, the first CA-MRSA strain isolated in Western Australia (WA), WA-5, encoded cadmium, tetracycline, and penicillin resistance genes on plasmid pWBG753 (∼30 kb). WA-5 and pWBG753 appeared only briefly in WA; however, fusidic acid resistance plasmids related to pWBG753 were also present in the first European CA-MRSA isolates at the time. Here, we characterize a 72-kb conjugative plasmid, pWBG731, present in multiresistant WA-5-like clones from the same period. pWBG731 was a cointegrant formed from pWBG753 and a pWBG749 family conjugative plasmid. pWBG731 carried mupirocin, trimethoprim, cadmium, and penicillin resistance genes. The stepwise evolution of pWBG731 likely occurred through the combined actions of IS, IS-dependent miniature inverted-repeat transposable elements (MITEs), and the BinL resolution system of the β-lactamase transposon Tn An evolutionarily intermediate ∼42-kb nonconjugative plasmid, pWBG715, possessed the same resistance genes as pWBG731 but retained an integrated copy of the small tetracycline resistance plasmid pT181. IS likely facilitated the replacement of pT181 with conjugation genes on pWBG731, thus enabling autonomous transfer. Like conjugative plasmid pWBG749, pWBG731 also mobilized nonconjugative plasmids carrying mimics. It seems likely that pWBG731 represents the product of multiple recombination events between the WA-5 pWBG753 plasmid and other mobile genetic elements present in indigenous community-associated methicillin-sensitive (CA-MSSA) isolates. The molecular evolution of pWBG731 saliently illustrates how diverse mobile genetic elements can together facilitate rapid accrual and horizontal dissemination of multiresistance in CA-MRSA.
