ErmE methyltransferase recognizes features of the primary and secondary structure in a motif within domain V of 23 S rRNA.
The Erm methyltransferases confer resistance to macrolide, lincosamide and streptogramin B (MLS) antibiotics by methylation of a single adenosine base within bacterial 23 S ribosomal RNA. The ErmE methyltransferase, from the macrolide-producing bacterium Saccharopolyspora erythraea, recognizes a motif within domain V of the rRNA that specifically targets adenosine 2058 (A2058) for methylation. Here, we define the structure of the RNA motif by a combination of molecular genetics and biochemical probing. The core of the motif has the primary sequence 2056-GGAHA-2060, where H is any nucleotide except guanosine, and ErmE methylates at the adenosine in bold. For efficient recognition by ErmE, this sequence must be displayed within a particular secondary structure. An irregular stem (helix 73) is required immediately 5' to A2058, with an unpaired nucleotide, preferably a cytidine residue, at position 2055. Nucleotides 2611 to 2616 are collectively required to form part of the 3'-side of helix 73, but there is little or no restriction on the identities of individual nucleotides here. There are minor preferences in the identities of nucleotides 2051 to 2055 that are adjacent to the motif core, although their main role is in maintaining the irregular secondary structure. The essential elements of the ErmE motif are conserved in bacterial 23 S rRNAs, and thus presumably also form the recognition motif for other Erm methyltransferases.
Villsen
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Department of Molecular Biology, Odense University, Campusvej 55, Odense M, DK-5230, Denmark.
Vester
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B
Douthwaite
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D000241:Adenosine / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism*; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000737:chemistry; D018942:Macrolides; D008745:Methylation; D008780:Methyltransferases / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D017354:Point Mutation; D012329:RNA, Bacterial / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects; D013379:Substrate Specificity