title: Heterologous Expression and Functional Characterization of the Exogenously Acquired Aminoglycoside Resistance Methyltransferases RmtD, RmtD2, and RmtG.
abstract: 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.
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA Laboratório de Investigação em Microbiologia Médica, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Laís L
authors.initials: LL
authors.lastname: Corrêa
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Marta A
authors.initials: MA
authors.lastname: Witek
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Natalia
authors.initials: N
authors.lastname: Zelinskaya
authors.affiliation: Laboratório de Investigação em Microbiologia Médica, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Renata C
authors.initials: RC
authors.lastname: Picão
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA gconn@emory.edu.
authors.forename: Graeme L
authors.identifier: http://orcid.org/0000-0002-2419-380X
authors.initials: GL
authors.lastname: Conn
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012435:S-Adenosylhomocysteine / Q000737:chemistry* / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D019076:Transgenes
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D007527:Isoenzymes; D011994:Recombinant Proteins; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; C446079:16S rRNA - m7G methyltransferase; D008780:Methyltransferases