Heterologous Expression and Functional Characterization of the Exogenously Acquired Aminoglycoside Resistance Methyltransferases RmtD, RmtD2, and RmtG

• Type of Material: Article
• Authors: Laís L. Corrêa, Emory University; Marta A. Witek, Emory University; Natalia Zelinskaya, Emory University; Renata C. Picão, Universidade Federal do Rio de Janeiro; Graeme Conn, Emory University
• Language: English
• Date: 2016-01-01
• Publisher: American Society for Microbiology
• Publication Version: Final Published Version
• Copyright Statement: Copyright © 2015, American Society for Microbiology. All Rights Reserved.
• Final Published Version (URL): http://dx.doi.org/10.1128/AAC.02482-15
• Title of Journal or Parent Work: Antimicrobial Agents and Chemotherapy
• ISSN: 0066-4804
• Volume: 60
• Issue: 1
• Start Page: 699
• End Page: 702
• Grant/Funding Information: National Institute of Allergy and Infectious Diseases (NIAID) provided funding to Marta A. Witek, Natalia Zelinskaya, and Graeme L. Conn under grant number R01-AI088025.; Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) provided funding to Renata C. Picão and Laís L. Corrêa under grant numbers E-26/111.780/2012 and E-26/ 201.555/2014.; MCTI; NIH; HHS; L.L.C. received support from a Ciência sem Fronteiras fellowship (CNPq).
• Keywords: KLEBSIELLA-PNEUMONIAE; ISOLATE; 16S RIBOSOMAL-RNA; COPRODUCTION; METHYLATION; SGM; Life Sciences & Biomedicine; Microbiology; ANTIBIOTICS; Pharmacology & Pharmacy; HIGH-LEVEL RESISTANCE
• Research Categories: Biology, Microbiology; Chemistry, Biochemistry; Health Sciences, Pharmacology

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 m7G1405 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 (m7G1405) methyltransferase family.