Molecular mechanism of trans-translation reaction
| Project/Area Number |
14360044
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Hirosaki University |
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Principal Investigator |
HIMENO Hyouta Hirosaki University, Faculty of Agriculture and Life Science, Associate Professor, 農学生命科学部, 助教授 (80208785)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2004: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2003: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2002: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | tmRNA / trans-translation / SmpB / translation / ribosome / aminoglycoside / tmRNA / パロモマイシン / アミノアシル化 |
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| Research Abstract |
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.
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Report
(4 results)
Research Products
(30 results)
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[Journal Article] A novel GTPase activated by the small subunit of ribosome.2004
Author(s)
Himeno, H., Hanawa-Suetsugu, K., Kimura, T., Takagi, K., Sugiyama, W., Shirata, S., Mikami, T., Odagiri, F., Osanai, Y., Watanabe, D., Goto, S., Kalachnyuk, L., Ushida, C., Muto, A.
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Journal Title
Nucleic Acids Res. 32
Pages: 5303-5309
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] In vitro analysis of the initial steps of trans-translation.2003
Author(s)
Takada, K., Hori-Takemoto, C., Kawazoe, M., Lee, S., Shirouzu, M., Yokoyama, S., Muto, A., Himeno, H.
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Journal Title
Nucleic Acids Res.Supplement 3
Pages: 287-288
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] In vitro analysis of the initial steps of trans-translation.2003
Author(s)
Takada, K., Hori-Takemoto, C., Kawazoe, M., Lee, S., Shirouzu, M., Yokoyama, S., Muto, A., Himeno, H.
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Journal Title
Nucleic Acids Res.Sup. 3
Pages: 287-288
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Detection of tmRNA-mediated trans-translation products in Bacillus subtilis.2002
Author(s)
Fujihara, A., Tomatsu, H., Inagaki, S., Tadaki, T., Ushida, C., Himeno, H., Muto, A.
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Journal Title
Genes to Cells 7
Pages: 343-350
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] Trans-translation mediated by Bacillus subtilis tmRNA.2002
Author(s)
Ito, K., Tadaki, T., Lee, S., Takada, K., Muto, A., Himeno, H.
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Journal Title
FEBS Lett. 516
Pages: 245-252
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] Trans-translation mediated by Bacillus subtilis tmRNA.2002
Author(s)
Ito, K., Tadaki, T., Lee, S., Takada, K., Mute, A., Himeno, H.
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Journal Title
FEBS Lett. 516
Pages: 245-252
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Publications] Takada, K., Hori-Takemoto, C., Kawazoe, M., Lee, S., Shirouzu, M., Yokoyama, S., Muto, A., Himeno, H.: "In vitro analysis of the initial steps of trans-translation"Nucleic Acids Res. Supplement. 3. 287-288 (2003)
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[Publications] Fujihara, A., Tomatsu, H., Inagaki, S., Tadaki, T., Ushida, C., Himeno, H., Muto, A.: "Detection of tmRNA-mediated trans-translation products in Bacillus subtilis"Genes to Cells. 7. 343-350 (2002)
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[Publications] Ito, K., Tadaki, T., Lee, S., Takada, K., Muto, A., Himeno, H.: "Trans-translation mediated by Bacillus subtilis tmRNA"FEBS Lett.. 516. 245-252 (2002)
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