EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance

Paul A. Mann, Liqun Xiong, Alexander S. Mankin, Andrew S. Chau, Cara A. Mendrick, David J. Najarian, Christina A. Cramer, David Loebenberg, Elizabeth Coates, Nicholas J. Murgolo, Frank M. Aarestrup, Richard V. Goering, Todd A. Black, Roberta S. Hare, Paul M. McNicholas

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Enterococcus faecium strain 9631355 was isolated from animal sources on the basis of its resistance to the growth promotant avilamycin. The strain also exhibited high-level resistance to evernimicin, a drug undergoing evaluation as a therapeutic agent in humans. Ribosomes from strain 9631355 exhibited a dramatic reduction in evernimicin binding, shown by both cell-free translation assays and direct-binding assays. The resistance determinant was cloned from strain 9631355; sequence alignments suggested it was a methyltransferase and therefore it was designated emtA for evernimicin methyltransferase. Evernimicin resistance was transmissible and emtA was localized to a plasmid-borne insertion element. Purified EmtA methylated 50S subunits from an evernimicin-sensitive strain 30-fold more efficiently than those from a resistant strain. Reverse transcription identified a pause site that was unique to the 23S rRNA extracted from resistant ribosomes. The pause corresponded to methylation of residue G2470 (Escherichia coli numbering). RNA footprinting revealed that G2470 is located within the evernimicin-binding site on the ribosome, thus providing an explanation for the reduced binding of the drug to methylated ribosomes.

Original languageEnglish
Pages (from-to)1349-1356
Number of pages8
JournalMolecular Microbiology
Volume41
Issue number6
DOIs
StatePublished - 2001

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Methyltransferases
Ribosomes
Enterococcus faecium
Drug Evaluation
Sequence Alignment
Methylation
Reverse Transcription
evernimicin
Plasmids
Binding Sites
RNA
Escherichia coli
Growth
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Microbiology

Cite this

Mann, P. A., Xiong, L., Mankin, A. S., Chau, A. S., Mendrick, C. A., Najarian, D. J., ... McNicholas, P. M. (2001). EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance. Molecular Microbiology, 41(6), 1349-1356. https://doi.org/10.1046/j.1365-2958.2001.02602.x

EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance. / Mann, Paul A.; Xiong, Liqun; Mankin, Alexander S.; Chau, Andrew S.; Mendrick, Cara A.; Najarian, David J.; Cramer, Christina A.; Loebenberg, David; Coates, Elizabeth; Murgolo, Nicholas J.; Aarestrup, Frank M.; Goering, Richard V.; Black, Todd A.; Hare, Roberta S.; McNicholas, Paul M.

In: Molecular Microbiology, Vol. 41, No. 6, 2001, p. 1349-1356.

Research output: Contribution to journalArticle

Mann, PA, Xiong, L, Mankin, AS, Chau, AS, Mendrick, CA, Najarian, DJ, Cramer, CA, Loebenberg, D, Coates, E, Murgolo, NJ, Aarestrup, FM, Goering, RV, Black, TA, Hare, RS & McNicholas, PM 2001, 'EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance', Molecular Microbiology, vol. 41, no. 6, pp. 1349-1356. https://doi.org/10.1046/j.1365-2958.2001.02602.x
Mann, Paul A. ; Xiong, Liqun ; Mankin, Alexander S. ; Chau, Andrew S. ; Mendrick, Cara A. ; Najarian, David J. ; Cramer, Christina A. ; Loebenberg, David ; Coates, Elizabeth ; Murgolo, Nicholas J. ; Aarestrup, Frank M. ; Goering, Richard V. ; Black, Todd A. ; Hare, Roberta S. ; McNicholas, Paul M. / EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance. In: Molecular Microbiology. 2001 ; Vol. 41, No. 6. pp. 1349-1356.
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