Abstract
Negamycin (NEG) is a ribosome-targeting antibiotic that exhibits clinically promising activity. Its binding site and mode of action have remained unknown. We solved the structure of the Thermus thermophilus ribosome bound to mRNA and three tRNAs, in complex with NEG. The drug binds to both small and large ribosomal subunits at nine independent sites. Resistance mutations in the 16S rRNA unequivocally identified the binding site in the vicinity of the conserved helix 34 (h34) in the small subunit as theprimary site of antibiotic action in the bacterial and, possibly, eukaryotic ribosome. At this site, NEG contacts 16S rRNA as well as the anticodon loop of the A-site tRNA. Although the NEG site of action overlaps with that of tetracycline (TET), the two antibiotics exhibit different activities: while TET sterically hinders binding of aminoacyl-tRNA to the ribosome, NEG stabilizes its binding, thereby inhibiting translocation and stimulating miscoding.
| Original language | English |
|---|---|
| Pages (from-to) | 541-550 |
| Number of pages | 10 |
| Journal | Molecular Cell |
| Volume | 56 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Nov 20 2014 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology
Cite this
Negamycin Interferes with Decoding and Translocation by Simultaneous Interaction with rRNA and tRNA. / Polikanov, Yury S.; Szal, Teresa; Jiang, Fuyan; Gupta, Pulkit; Matsuda, Ryoichi; Shiozuka, Masataka; Steitz, Thomas A.; Vázquez-Laslop, Nora; Mankin, Alexander S.
In: Molecular Cell, Vol. 56, No. 4, 20.11.2014, p. 541-550.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Negamycin Interferes with Decoding and Translocation by Simultaneous Interaction with rRNA and tRNA
AU - Polikanov, Yury S.
AU - Szal, Teresa
AU - Jiang, Fuyan
AU - Gupta, Pulkit
AU - Matsuda, Ryoichi
AU - Shiozuka, Masataka
AU - Steitz, Thomas A.
AU - Vázquez-Laslop, Nora
AU - Mankin, Alexander S.
PY - 2014/11/20
Y1 - 2014/11/20
N2 - Negamycin (NEG) is a ribosome-targeting antibiotic that exhibits clinically promising activity. Its binding site and mode of action have remained unknown. We solved the structure of the Thermus thermophilus ribosome bound to mRNA and three tRNAs, in complex with NEG. The drug binds to both small and large ribosomal subunits at nine independent sites. Resistance mutations in the 16S rRNA unequivocally identified the binding site in the vicinity of the conserved helix 34 (h34) in the small subunit as theprimary site of antibiotic action in the bacterial and, possibly, eukaryotic ribosome. At this site, NEG contacts 16S rRNA as well as the anticodon loop of the A-site tRNA. Although the NEG site of action overlaps with that of tetracycline (TET), the two antibiotics exhibit different activities: while TET sterically hinders binding of aminoacyl-tRNA to the ribosome, NEG stabilizes its binding, thereby inhibiting translocation and stimulating miscoding.
AB - Negamycin (NEG) is a ribosome-targeting antibiotic that exhibits clinically promising activity. Its binding site and mode of action have remained unknown. We solved the structure of the Thermus thermophilus ribosome bound to mRNA and three tRNAs, in complex with NEG. The drug binds to both small and large ribosomal subunits at nine independent sites. Resistance mutations in the 16S rRNA unequivocally identified the binding site in the vicinity of the conserved helix 34 (h34) in the small subunit as theprimary site of antibiotic action in the bacterial and, possibly, eukaryotic ribosome. At this site, NEG contacts 16S rRNA as well as the anticodon loop of the A-site tRNA. Although the NEG site of action overlaps with that of tetracycline (TET), the two antibiotics exhibit different activities: while TET sterically hinders binding of aminoacyl-tRNA to the ribosome, NEG stabilizes its binding, thereby inhibiting translocation and stimulating miscoding.
UR - http://www.scopus.com/inward/record.url?scp=84912565442&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84912565442&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2014.09.021
DO - 10.1016/j.molcel.2014.09.021
M3 - Article
C2 - 25306922
AN - SCOPUS:84912565442
VL - 56
SP - 541
EP - 550
JO - Molecular Cell
JF - Molecular Cell
SN - 1097-2765
IS - 4
ER -