Research Project
Grant-in-Aid for Scientific Research (B)
Trans-translation is an unusual translation in which tmRNA plays a dual function as a tRNA and an mRNA to relieve the stalled translation on the ribosome. In this study, we examined the effects of several kinds of aminoglycoside antibiotics, on trans-translation in vitro. A chemical footprinting study revealed that paromomycins bind tmRNA in the tRNA domain and in the middle of the long helix between tRNA and mRNA domains. Paromomycin bound in the tRNA domain inhibited aminoacylation, and the inhibition was suppressed by the addition of SmpB, a tmRNA binding protein. It was also found that paromomycin causes a shift of the translation-resuming point on tmRNA by -1. The effect on initiation-shift was suppressed by a mutation at the paromomycin binding site in 16S rRNA, but not by mutations in tmRNA. The effect of paromomycin on transtranslation differs substantially from that on canonical translation, in which it induces miscoding by modulating the A site of the decoding helix of the small subunit RNA of the ribosome. A minimal structure that causes initiation-shift was identified. The effect of each aminoglycoside on trans-translation coincides with that on conformational change in the A site of the small subunit of the ribosome revealed by recent structural studies : paromomycin, tobramycin and genetic in that is categorized into the gentamicin subclass, but not streptomycin and hygromycin B, flip out two conserved adenine bases at 1492 and 1493 from the A site helix.
All 2004 2003 2002 Other
All Journal Article (18 results) Book (2 results) Publications (10 results)
細胞における蛋白質の一生(蛋白質核酸酵素増刊) 49(7)
Pages: 1062-1066
Nucleic Acids Res. 32
Pages: 4119-4126
Pages: 5303-5309
Nucleic Acids Symp.Ser. 48
Pages: 299-300
Life of Proteins : Synthesis, Maturation : Traffic, Quality Control, Degradation, Disease (Protein, Nucleic Acid and Enzyme suppl) 49(7)
J.Biol.Chem. 278
Pages: 27672-27680
Nucleic Acids Res.Supplement 3
Pages: 236-236
Pages: 287-288
Nucleic Acids Res.Sup. 3
Pages: 235-236
Genes to Cells 7
Pages: 343-350
Nucleic Acids Res. 30
Pages: 1620-1629
FEBS Lett. 516
Pages: 245-252
RNAの細胞生物学(蛋白質核酸酵素増刊) 48(4)
Pages: 346-356
RNA Biology of the Cell (Protein, Nucleic Acid and Enzyme suppl) 48(4)