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The mecA gene, the central determinant of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA), is not native to this bacterial species but may have originated in the animal commensal species Staphylococcus sciuri. All S. sciuri strains carry a close homologue of mecA in the form of pbpD, the genetic determinant of penicillin binding protein 4 (PBP 4) of S. sciuri. Here we describe an experimental system that could be used for additional tests for this proposition. The S. sciuri pbpD gene was cloned into a shuttle plasmid and introduced into methicillin-susceptible S. aureus strain COL-S derived from parental MRSA strain COL from which the resistance cassette staphylococcal cassette chromosome mec was excised. The S. sciuri pbpD determinant was transcribed and translated in the S. aureus transductants producing large amounts of the 84-kDa S. sciuri PBP 4 and was then deposited in the plasma membrane of the host bacterium. Transductants carrying the heterologous S. sciuri pbpD gene exhibited properties typical of those of parental MRSA strain COL, including broad-spectrum, high-level, and homogeneous resistance to structurally different beta-lactams. Antibiotic resistance was dependent on the functioning of S. aureus PBP 2 and was suppressed by the specific regulatory genes mecI and mecR and by inhibitors of an early step in cell wall biosynthesis. S. sciuri PBP 4 was also able to replace the essential physiological function(s) of the native PBP 2 of S. aureus and produce peptidoglycan typical of that of parental MRSA strain COL. Our results provide further support for the proposition that the resistance determinant mecA of MRSA strains has evolved from S. sciuri pbpD.

antimicrobial agents and chemotherapy

Reconstruction of the Phenotypes of Methicillin-Resistant Staphylococcus aureu s by Replacement of the Staphylococcal Cassette Chromosome mec with a Plasmid-Borne Copy of Staphylococcus sciuri pbpD Gene