Molecular mechanism of trans-translation mediated by tmRNA and its application
Project/Area Number |
17380061
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Applied biochemistry
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Research Institution | Hirosaki University |
Principal Investigator |
MUTO Akira Hirosaki University, Faculty of Biochemistry and Life Science, Research Scientist (80034635)
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Co-Investigator(Kenkyū-buntansha) |
姫野 俵太 弘前大学, 農学生命科学部, 教授 (80208785)
USHIDA Chisato Hirosaki University, Faculty of Biochemistry and Life Science, professor (50250593)
|
Project Period (FY) |
2005 – 2007
|
Project Status |
Completed (Fiscal Year 2007)
|
Budget Amount *help |
¥15,890,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2007: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2006: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2005: ¥5,700,000 (Direct Cost: ¥5,700,000)
|
Keywords | tmRNA / trans-translation / Bacillus subtlis / SmpB / ribosome / Stress response / RNA / Escherichia coli / 胞子形成 / 分子擬態 / タグペプチド |
Research Abstract |
a) Molecular mechanism of trans-translation mediated by tmRNA To investigate the functional structure of tmRNA in trans-translation reaction, we have developed an in vitro trans-translation system using E. coli extracts. Many mutant tmRNAs were constructed by in vitro mutagenesis, and tested their activities as tRNA, mRNA and rribosome binding. From the results, we could identify the structures and sequences in tmRNA required for each function. The effect of several antibiotics on the trans-translation was investigated: we found that paromomycin directly binds to tmRNA causing the frame-shift of translation of tag-peptides. b) Physiological roles of trans-translation We have constructed an in vivo system in which trans-translation products can be detected in Bacillus subtilis and E. coli. Using this system, several reaction products were isolated and identified by the N-terminal amino acid sequence analyses. The sites of trans-translation in some of the products were determined by MAS-spectrometry. It was shown that the trans-translation is involved in various cellular activities, such as catabolite repression, amino acid metabolism, stress responses, sporulation and so on. Endspore formation of Bacillus subtilis strain is significantly impaired by deleting the gene for tmRNA. The genetic and biochemical experimental results revealed that the synthesis of sigma-Kduring the late stage of sporulation process is significantly impaired in the mutant cells.
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Report
(4 results)
Research Products
(69 results)
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[Journal Article] In vitro trans-translation of Thermus thermophilus: Ribosomal protein S1 is not required for the early stage of trans-translation. A2007
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Hanawa-Suetsugu, K., Lee, S, Shirouzu, M., Yokoyama, S., Muto, A. & Himeno, H
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Journal Title
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
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[Journal Article] In vitro trans-translation of Thermus thermophilus: Ribosomal protein S1 is not required for the early stage of trans-translation2007
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Hanawa-Suetsugu, K., Lee, S, Shirouzu, M., Yokoyama, S., Muto, A. & Himeno, H
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Journal Title
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Molecular mechanism of trans-translation2007
Author(s)
Himeno, H., Kurita, D., Tatada, K., Konno, T., Hanawa, K., Takemoto, C., Kawazoe, M., Yokoyama, S., Nameki, N., Kawi, G & Muto, A.
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Journal Title
Biochemistry (Tokyo) 79
Pages: 213-221
NAID
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] In vitro trans-translation of Thermus thermophilus : Ribosomal protein S1 is not required for the early stage of trans-translation.2007
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Hanawa-Suetsugu, K., Lee, S., Shirouzu, M., Yokoyama, S., Muto, A., Himeno, H.
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Journal Title
Related Report
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[Journal Article] トランス・トランスレーションの分子メカニズム2007
Author(s)
姫野俵太, 栗田大輔, 高田一馬, 今野貴之, 塙(末次)京子, 竹本千重, 川添将仁, 横山茂之, 行木信一, 河合剛太, 武藤あきら
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[Journal Article] Twelve novel C. elegans RNA candidates isolated by two-dimensional polyacrylamide gel electrophoresis. Gene 365 (2006).2006
Author(s)
Hokii, Y., Kubo, A., Ogasawara, T., Nogi, Y., Taneda, A., Arai, R., Muto, A., Ushida, C.
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Journal Title
Description
「研究成果報告書概要(和文)」より
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Peer Reviewed
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[Journal Article] Twelve novel C. elegans RNA candidates isolated by two-dimensional polyacrylamide gel electrophoresis2006
Author(s)
Hokii, Y., Kubo, A., Ogasawara, T., Nogi, Y., Taneda, A., Arai, R., Muto, A., Ushida, C.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] An RNA polymerase inhibitor, cyclothiazomycin B1, and its isomer.2006
Author(s)
Hashimoto, M., Murakami, T., Funahashi, K., Tokunaga, T., Nihei, K., Okuno, T., Kimura, T., Himeno, H.
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Journal Title
Bioorg. Med. Chem. 14
Pages: 8259-8270
Related Report
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[Journal Article] Twelve novel C elegans RNA candidates isolated by two-dimensional polyacrylamide gel electrophoresis2006
Author(s)
Hokii, Y., Kubo, A., Ogasawara, T., Nogi, Y., Tanada, A., Arai, R., Muto, A., Ushida, C.
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Journal Title
Related Report
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[Journal Article] Competition between trans-translation and termination or elongation of translation2005
Author(s)
Asano, K., Kurita, D., Takada, K., Konno, T., Muto, A. & Himeno, H.
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Journal Title
Nucleic Acids Res. 33
Pages: 5544-5552
Description
「研究成果報告書概要(和文)」より
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Peer Reviewed
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[Journal Article] Interaction analysis between tmRNA and SmpB from Thermus thermophilus2005
Author(s)
Nameki, N., Someya, T., Okano, S., Suemasa, R., Kimoto, M., Hanawa-Suetsugu, K., Terada, T., Shirouzu, M., Hirao, I, Takaku, H., Himeno, H., Muto, A., Kuramitsu, S., Yokoyama, S. & Kawai, G.
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Journal Title
J. Biochem. 138
Pages: 729-739
NAID
Description
「研究成果報告書概要(和文)」より
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Peer Reviewed
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[Journal Article] Interaction analysis between tmRNA and SmpB from Thermus thermophilus2005
Author(s)
Nameki, N., Someya, T., Okano, S., Suemasa, R., Kimoto, M., Hanawa-Suetsugu, K., Terada, T., Shirouzu, M., Hirao, I., Takaku, H., Himeno, H., Muto, A., Kuramitsu, S., Yokoyama, S. & Kawai, G.
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Journal Title
J. Biochem. 138
Pages: 729-739
NAID
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Trans-translation: in "Life of Protein in the Cell-Synthesis, Maturation, Transportation, Control, Degradation and Disease-" (in Japanese) (Ogura, H., Endo, T., Mori, M, & Yoshida, T. eds.)2005
Author(s)
Himeno, H. & Muto., A.
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Journal Title
Kyoritu Shuppan
Pages: 1062-1066
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Functional analysis of factors involved in trans-translation2005
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Matsuda, N., Kurita, D., Asano, K., Shirouzu, M., Yokoyama, S., Muto, A., Himeno, H.
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Journal Title
Nucleic Acids Symp.Ser 49
Pages: 101-102
Related Report
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[Journal Article] Competition between trans-trnslation and termination or elongation of translation2005
Author(s)
Asano, K., Kurita, D., Takada, K., Konno, T., Muto, A., Himeno, H.
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Journal Title
Nucleic Acids Res. 33
Pages: 5544-5552
Related Report
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[Presentation] Search for tRNA genes: construction of tRNA gene data-base2007
Author(s)
Kohara, Y., Inokuchi, H., Muto, A., Yamada, Y., Kinouchi, M., Maeno, T., Kanaya, S., Ikeura, T. & Abe, T.
Organizer
The annual meeting of Molecular Biology and Biochemistry 2007
Place of Presentation
Yokohama
Year and Date
2007-12-13
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Trans-translation mediated by tmRNA and SmpB2007
Author(s)
Kurita, D., Konno, T., Takada, K., Asano, K., Muto, A. & Himeno, H.
Organizer
RNA 2006 Izu"Functional RNAs and Regulatory Machinery"
Place of Presentation
伊豆
Year and Date
2007-12-05
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Trans-translation mediated by tmRNA and SmpB2007
Author(s)
Kurita, D., Konno, T., Takada, K., Asano, K., Muto, A. & Himeno, H.
Organizer
RNA 2006, Izu "Functional RNAs and Regulatory Machinery"
Place of Presentation
Izu, Japan
Year and Date
2007-12-05
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Trans-translation mediated by tmRNA and SmpB2007
Author(s)
Kurita, D., Konno, T., Takada, K., Asano, K., Muto, A. & Himeno, H.
Organizer
RNA 2006 Izu Functional RNAs and Regulatory Machinery
Place of Presentation
伊豆
Year and Date
2007-12-05
Related Report
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[Presentation] Ribosomal rotein S1 is not involved in the early stage of trans-translation2007
Author(s)
Tatada, K., Kurita, D., Konno, T., Hanawa, K., Takemoto, C., Kawazoe, M., Yokoyama, S., Nameki, N., Kawi, G., Muto., A. & Himeno, H.
Organizer
The 9th RNA meeting
Place of Presentation
Nagoya
Year and Date
2007-07-29
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Expression of C. elegans novel ncRNAs. Regularoty RNAs2006
Author(s)
Ushida, C., Hokii, Y., Sugawara, Y., Mizuki, Y., Endo, Y. & Muto, A.
Organizer
The LXXI Cold Spring Harbor Sympoium
Place of Presentation
Cold Spring Harbor, USA
Year and Date
2006-07-20
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Expression of C. elegans novel ncRNAs: Regularoty RNAs2006
Author(s)
Ushida, C., Hokii, Y., Sugawara, Y., Mizuki, Y., Endo, Y. & Muto, A.
Organizer
The LXXI Cold Spring Harbor Symposium
Place of Presentation
Cold Spring Harbor, USA
Year and Date
2006-07-20
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] 「研究成果報告書概要(欧文)」より2006
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Asano, K., Sirouzu, M., Yokoyama, S., Muto, A. & Himeno, H.
Organizer
The 8th RNA meeting
Place of Presentation
Awaji
Year and Date
2006-07-18
Related Report
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[Presentation] Inhibition of GTPase activity of RsgA by novel antibiotics2006
Author(s)
Kimura, T., Murakami, T., Hashimoto, M., Okuno, T., Muto, A., & Himeno, H.
Organizer
The 8th RNA meeting
Place of Presentation
Awaji
Year and Date
2006-07-18
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Interaction of SmpB with ribosome by hydroxyradical footprinting2006
Author(s)
Kimura, T., Konno, T., Sasaki, R., Asano, K., Takada, K., Muto, A. & Himeno, H.
Organizer
The 8th RNA meeting
Place of Presentation
Awaji
Year and Date
2006-07-18
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Effect of Cyclothazomycin B1 on a novel GTPase activated by small ribosomal subunit, RsgA2006
Author(s)
Kimura, T., Murakami, T., Hashimoto, M., Okuno, T., Muto, A., & Himeno, H.
Organizer
The annual meeting of Agricultural Biochemistry
Place of Presentation
Kyoto
Year and Date
2006-03-26
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Functional analysis of factors involved in trans-translation. (2005)2005
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Matsuta, N., Kurita, D., Shirouzu, M., Yokoyama, S., Muto, A. & Himeno, H.
Organizer
The 4th International Symposium on Nucleic Acids Chemistry
Place of Presentation
東京
Year and Date
2005-10-20
Description
「研究成果報告書概要(和文)」より
Related Report
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[Presentation] Functional analysis of factors involved in trans-translation2005
Author(s)
Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Matsuta, N., Kurita, D., Shirouzu, M., Yokoyama, S., Muto, A. & Himeno, H.
Organizer
The 4th International Symposium on Nucleic Acids Chemistry
Place of Presentation
Tokyo, Japan
Year and Date
2005-10-20
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] The shift of trans-translation initiation by aminoglycosides2005
Author(s)
Konno, T., Takahashi, T., Kurita, D., Asano, K., Miura, H., Muto, A. & Himeno, H.
Organizer
The 7th RNA meeting
Place of Presentation
Hirosaki
Year and Date
2005-08-10
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Competitions between tmRNA, tRNA, RF and RRF on the A-ste of stalled ribosome2005
Author(s)
Asano, K., Konno, T., Kurita, D., Miura, H., Muto, A. & Himeno, H.
Organizer
The 7th RNA meeting
Place of Presentation
Hirosaki
Year and Date
2005-08-10
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Presentation] Involvement of mRNA-mediated trans-translation in sporulation process of Bacillus subtilis2005
Author(s)
Abe, T., Sakaki, T., Fujihara, A., Otsuka, H., Ujiie, H., Ushida, C., Himeno, H. & Sato, T. & Muto, A.
Organizer
The 7th RNA meeting
Place of Presentation
Hirosaki
Year and Date
2005-08-10
Description
「研究成果報告書概要(欧文)」より
Related Report
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