title: The order Bacillales hosts functional homologs of the worrisome cfr antibiotic resistance gene.
abstract: The cfr gene encodes the Cfr methyltransferase that methylates a single adenine in the peptidyl transferase region of bacterial ribosomes. The methylation provides resistance to several classes of antibiotics that include drugs of clinical and veterinary importance. This paper describes a first step toward elucidating natural residences of the worrisome cfr gene and functionally similar genes. Three cfr-like genes from the order Bacillales were identified from BLAST searches and cloned into plasmids under the control of an inducible promoter. Expression of the genes was induced in Escherichia coli, and MICs for selected antibiotics indicate that the cfr-like genes confer resistance to PhLOPSa (phenicol, lincosamide, oxazolidinone, pleuromutilin, and streptogramin A) antibiotics in the same way as the cfr gene. In addition, modification at A2503 on 23S rRNA was confirmed by primer extension. Finally, expression of the Cfr-like proteins was verified by SDS gel electrophoresis of whole-cell extracts. The work shows that cfr-like genes exist in the environment and that Bacillales are natural residences of cfr-like genes.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark.
authors.forename: Lykke H
authors.initials: LH
authors.lastname: Hansen
authors.forename: Mercè H
authors.initials: MH
authors.lastname: Planellas
authors.forename: Katherine S
authors.initials: KS
authors.lastname: Long
authors.forename: Birte
authors.initials: B
authors.lastname: Vester
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D056449:Bacillales / Q000187:drug effects* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004224:Diterpenes / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects; D055231:Lincosamides / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D023303:Oxazolidinones / Q000494:pharmacology; D011083:Polycyclic Compounds; D025364:Streptogramin A / Q000494:pharmacology; D000097567:Pleuromutilins
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004224:Diterpenes; D055231:Lincosamides; D023303:Oxazolidinones; D011083:Polycyclic Compounds; D025364:Streptogramin A; D000097567:Pleuromutilins