Negamycin Interferes with Decoding and Translocation by Simultaneous Interaction with rRNA and tRNA

Yury S. Polikanov, Teresa Szal, Fuyan Jiang, Pulkit Gupta, Ryoichi Matsuda, Masataka Shiozuka, Thomas A. Steitz, Nora Vázquez-Laslop, Alexander S. Mankin

Research output: Contribution to journalArticle

23 Citations (Scopus)

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 languageEnglish
Pages (from-to)541-550
Number of pages10
JournalMolecular Cell
Volume56
Issue number4
DOIs
Publication statusPublished - Nov 20 2014
Externally publishedYes

Fingerprint

Transfer RNA
Ribosomes
Anti-Bacterial Agents
Tetracycline
Binding Sites
Large Ribosome Subunits
Small Ribosome Subunits
Thermus thermophilus
Anticodon
negamycin
Messenger RNA
Mutation
Pharmaceutical Preparations

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 journalArticle

Polikanov, YS, Szal, T, Jiang, F, Gupta, P, Matsuda, R, Shiozuka, M, Steitz, TA, Vázquez-Laslop, N & Mankin, AS 2014, 'Negamycin Interferes with Decoding and Translocation by Simultaneous Interaction with rRNA and tRNA', Molecular Cell, vol. 56, no. 4, pp. 541-550. https://doi.org/10.1016/j.molcel.2014.09.021
Polikanov, Yury S. ; Szal, Teresa ; Jiang, Fuyan ; Gupta, Pulkit ; Matsuda, Ryoichi ; Shiozuka, Masataka ; Steitz, Thomas A. ; Vázquez-Laslop, Nora ; Mankin, Alexander S. / Negamycin Interferes with Decoding and Translocation by Simultaneous Interaction with rRNA and tRNA. In: Molecular Cell. 2014 ; Vol. 56, No. 4. pp. 541-550.
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