Molecular mechanism of trans-translation reaction

• Project/Area Number: 14360044
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 応用微生物学・応用生物化学
• Research Institution: Hirosaki University
• 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)
• Project Period (FY): 2002 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥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)
• Keywords: tmRNA / trans-translation / SmpB / translation / ribosome / aminoglycoside / tmRNA / パロモマイシン / アミノアシル化

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.

Research Products

• [Journal Article] 翻訳と共役した蛋白質品質管理システム:トランストランスレーション (2004)
  • Author(s): 姫野俵太, 武藤
  • Journal Title: 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
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A minimum structure of aminoglycosides that causes an initiation shift of trans-translation. (2004)
  • Author(s): Konno, T., Takahashi, T., Kurita, D., Muto, A., Himeno, H.
  • Journal Title: Nucleic Acids Res. 32 Pages: 4119-4126
  • Description: 「研究成果報告書概要(和文)」より
• [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.
  • Journal Title: Nucleic Acids Res. 32 Pages: 5303-5309
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Initiation shift of trans-translation by aminoglycosides. (2004)
  • Author(s): Konno, T., Kurita, D., Takahashi, T., Muto, A., Himeno, H.
  • Journal Title: Nucleic Acids Symp.Ser. 48 Pages: 299-300
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Trans-translation : Co-translational protein quality control system. (2004)
  • Author(s): Muto, A., Himeno, H.
  • Journal Title: Life of Proteins : Synthesis, Maturation : Traffic, Quality Control, Degradation, Disease (Protein, Nucleic Acid and Enzyme suppl) 49(7) Pages: 1062-1066
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 翻訳と共役した蛋白質品質管理システム:トランストランスレーション (2004)
  • Author(s): 姫野俵太, 武藤 あきら
  • Journal Title: 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
• [Journal Article] A minimum structure of aminoglycosides that causes an initiation shift of trans-translation. (2004)
  • Author(s): Konno, T., Takahashi, T., Kurita D., Muto, A., Himeno, H.
  • Journal Title: Nucleic Acids Res. 32 Pages: 4119-4126
• [Journal Article] Various effects of paromomycin on tmRNA-directed trans-translation. (2003)
  • Author(s): Takahashi, T., Konno, T., Muto, A., Himeno, H.
  • Journal Title: J.Biol.Chem. 278 Pages: 27672-27680
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Various effects of paromomycin on tmRNA-mediated trans-translation. (2003)
  • Author(s): Konno, T., Takahashi, T., Muto, A., Himeno, H.
  • Journal Title: Nucleic Acids Res.Supplement 3 Pages: 236-236
  • Description: 「研究成果報告書概要(和文)」より
• [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.
  • Journal Title: Nucleic Acids Res.Supplement 3 Pages: 287-288
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Various effects of paromomycin on tmRNA-mediated trans-translation. (2003)
  • Author(s): Konno, T., Takahashi, T., Muto, A., Himeno, H.
  • Journal Title: Nucleic Acids Res.Sup. 3 Pages: 235-236
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Nucleic Acids Res.Sup. 3 Pages: 287-288
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Genes to Cells 7 Pages: 343-350
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] SmpB functions in various steps of trans-translation. (2002)
  • Author(s): Hanawa-Suetsugu, K., Takagi, M., Inokuchi, H., Himeno, H., Muto, A.
  • Journal Title: Nucleic Acids Res. 30 Pages: 1620-1629
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Trans-translation mediated by Bacillus subtilis tmRNA. (2002)
  • Author(s): Ito, K., Tadaki, T., Lee, S., Takada, K., Muto, A., Himeno, H.
  • Journal Title: FEBS Lett. 516 Pages: 245-252
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] tmRNAによるトランス翻訳機構と生理的機能 (2002)
  • Author(s): 武藤, 姫野俵太
  • Journal Title: RNAの細胞生物学(蛋白質核酸酵素増刊) 48(4) Pages: 346-356
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Trans-translation mediated by Bacillus subtilis tmRNA. (2002)
  • Author(s): Ito, K., Tadaki, T., Lee, S., Takada, K., Mute, A., Himeno, H.
  • Journal Title: FEBS Lett. 516 Pages: 245-252
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular mechanism and physiological roles of trans-translation mediatedby tmRNA (2002)
  • Author(s): Muto, A., Himeno, H.
  • Journal Title: RNA Biology of the Cell (Protein, Nucleic Acid and Enzyme suppl) 48(4) Pages: 346-356
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] 命化学II-遺伝子の働きとその応用- (2003)
  • Author(s): 渡辺公綱, 姫野俵太
  • Total Pages: 255
  • Publisher: 丸善
  • Description: 「研究成果報告書概要(和文)」より
• [Book] Chemistry of life -Roles and application of the gene- (2003)
  • Author(s): Watanabe, K., Himeno, H.
  • Total Pages: 256
  • Publisher: Maruzen
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Takahashi, T., Konno, T., Muto, A., Himeno, H.: "Various effects of paromomycin on tmRNA-directed trans-translation"J.Biol.Chem.. 278. 27672-27680 (2003)
• [Publications] Konno, T., Takahashi, T., Muto, A., Himeno, H.: "Various effects of paromomycin on tmRNA-mediated trans-translation"Nucleic Acids Res. Supplement. 3. 235-236 (2003)
• [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)
• [Publications] 姫野俵太: "タンパク質がわかる"竹縄忠臣編(羊土社). 10 (2003)
• [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)
• [Publications] Hanawa-Suetsugu, K., Takagi, M., Inokuchi, H., Himeno, H., Muto, A.: "SmpB functions in various steps of trans-translation"Nucleic Acids Res.. 30. 1620-1629 (2002)
• [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)
• [Publications] Takada, K., Hanawa, K., Lee, S., Himeno, H., Muto, A.: "The structure and function of tmRNA"Nucleic Acids Res.Supplement. 2. 65-66 (2002)
• [Publications] 武藤, 姫野俵太: "tmRNAによるトランス翻訳と生理的機能"蛋白質核酸酵素. 48・4. 346-356 (2003)
• [Publications] 渡辺公綱, 姫野俵太: "生命化学II -遺伝子の働きとその応用-"丸善. 255 (2003)