RESUMEN

The exogenously acquired 16S rRNA methyltransferases RmtD, RmtD2, and RmtG were cloned and heterologously expressed in Escherichia coli, and the recombinant proteins were purified to near homogeneity. Each methyltransferase conferred an aminoglycoside resistance profile consistent with m(7)G1405 modification, and this activity was confirmed by in vitro 30S methylation assays. Analyses of protein structure and interaction with S-adenosyl-l-methionine suggest that the molecular mechanisms of substrate recognition and catalysis are conserved across the 16S rRNA (m(7)G1405) methyltransferase family.

Asunto(s)

Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana/genética , Escherichia coli/genética , Metiltransferasas/genética , S-Adenosilhomocisteína/química , S-Adenosilmetionina/química , Aminoglicósidos/farmacología , Antibacterianos/farmacología , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Clonación Molecular , Escherichia coli/efectos de los fármacos , Escherichia coli/enzimología , Expresión Génica , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Cinética , Klebsiella pneumoniae/enzimología , Klebsiella pneumoniae/genética , Metiltransferasas/química , Metiltransferasas/metabolismo , Pseudomonas aeruginosa/enzimología , Pseudomonas aeruginosa/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , S-Adenosilhomocisteína/metabolismo , S-Adenosilmetionina/metabolismo , Especificidad por Sustrato , Transgenes