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 language | English |
|---|---|
| Pages (from-to) | 1349-1356 |
| Number of pages | 8 |
| Journal | Molecular Microbiology |
| Volume | 41 |
| Issue number | 6 |
| DOIs | |
| State | Published - 2001 |
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All Science Journal Classification (ASJC) codes
- Molecular Biology
- Microbiology
Cite this
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 journal › Article
}
TY - JOUR
T1 - EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance
AU - Mann, Paul A.
AU - Xiong, Liqun
AU - Mankin, Alexander S.
AU - Chau, Andrew S.
AU - Mendrick, Cara A.
AU - Najarian, David J.
AU - Cramer, Christina A.
AU - Loebenberg, David
AU - Coates, Elizabeth
AU - Murgolo, Nicholas J.
AU - Aarestrup, Frank M.
AU - Goering, Richard V.
AU - Black, Todd A.
AU - Hare, Roberta S.
AU - McNicholas, Paul M.
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0034786898&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034786898&partnerID=8YFLogxK
U2 - 10.1046/j.1365-2958.2001.02602.x
DO - 10.1046/j.1365-2958.2001.02602.x
M3 - Article
C2 - 11580839
AN - SCOPUS:0034786898
VL - 41
SP - 1349
EP - 1356
JO - Molecular Microbiology
JF - Molecular Microbiology
SN - 0950-382X
IS - 6
ER -