Molecular mechanism of trans-translation mediated by tmRNA and its application

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17380061
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Hirosaki University
• Principal Investigator: MUTO Akira Hirosaki University, Faculty of Biochemistry and Life Science, Research Scientist (80034635)
• 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
• Keywords: tmRNA / trans-translation / Bacillus subtlis / SmpB / ribosome / Stress response / RNA / Escherichia coli

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.

Research Products

• [Journal Article] In vitro trans-translation of Thermus thermophilus: Ribosomal protein S1 is not required for the early stage of trans-translation. A (2007)
  • Author(s): Takada, K., Takemoto, C., Kawazoe, M., Konno, T., Hanawa-Suetsugu, K., Lee, S, Shirouzu, M., Yokoyama, S., Muto, A. & Himeno, H
  • Journal Title: RNA 13 Pages: 503-510
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A functional interaction of SmpB with tmRNA for determination of the resuming point of trans-translation (2007)
  • Author(s): Konno, T., Kurita, D., Takada, K., Muto, A. & Himeno, H.
  • Journal Title: RNA 13 Pages: 1723-1731
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Interaction of SmpB with ribosome from directed hydroxyl radical probing. 35 (2007) (2007)
  • Author(s): Kurita, D., Sasaki, R., Muto, A. & Himeno, H.
  • Journal Title: Nucleic Acids Res. 35 Pages: 7248-7255
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [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
  • Journal Title: 生化学 79 Pages: 213-221
  • Description: 「研究成果報告書概要(和文)」より
• [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
  • Journal Title: RNA 13 Pages: 503-510
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interaction of SmpB with ribosome from directed hydroxyl radical probing (2007)
  • Author(s): Kurita, D., Sasaki, R., Muto, A. & Himeno, H.
  • Journal Title: Nucleic Acids Res. 35 Pages: 7248-7255
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular mechanism of trans-translation (2007)
  • Author(s): Himeno, H., Kurita, D., Tatada, K., Konno, T., Hanawa, K., Takemoto, C., Kawazoe, M., Yokoyama, S., Nameki, N., Kawi, G & Muto, A.
  • Journal Title: Biochemistry (Tokyo) 79 Pages: 213-221
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Gene 365 Pages: 83-87
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] トランストランスレーション (2006)
  • Author(s): 姫野俵太、武藤あき, ら
  • Journal Title: 蛋白質核酸酵素 51 Pages: 2321-2327
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Twelve novel C. elegans RNA candidates isolated by two-dimensional polyacrylamide gel electrophoresis (2006)
  • Author(s): Hokii, Y., Kubo, A., Ogasawara, T., Nogi, Y., Taneda, A., Arai, R., Muto, A., Ushida, C.
  • Journal Title: Gene 365 Pages: 83-87
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trans-translation (2006)
  • Author(s): Himeno, H. & Muto., A.
  • Journal Title: Protein Nucl. Acid Enzyme 51 Pages: 2321-2327
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Competition between trans-translation and termination or elongation of translation (2005)
  • Author(s): Asano, K., Kurita, D., Takada, K., Konno, T., Muto, A. & Himeno, H.
  • Journal Title: Nucleic Acids Res. 33 Pages: 5544-5552
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Interaction analysis between tmRNA and SmpB from Thermus thermophilus (2005)
  • 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.
  • Journal Title: J. Biochem. 138 Pages: 729-739
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Participation of 3'-to-5' exoribonucleases in the turnover of Bacillus subtilis mRNA (2005)
  • Author(s): Oussenko, I. A., Abe, T., Ujiie, H., Muto, A. & Bechhofer, D.H.
  • Journal Title: J. Bacteriol 187 Pages: 2758-2767
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Participation of 3''-to-5'' Exoribonucleases in the turnover of Bacillus subtilis Messenger RNA (2005)
  • Author(s): Oussenko, I., Abe, T., Ujiie, H., Muto, A., Bechhofer, D. A.
  • Journal Title: J. Bacteriol. 187 Pages: 2758-2767
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interaction analysis between tmRNA and SmpB from Thermus thermophilus (2005)
  • 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.
  • Journal Title: J. Biochem. 138 Pages: 729-739
  • Description: 「研究成果報告書概要(欧文)」より
• [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.
  • Journal Title: Kyoritu Shuppan Pages: 1062-1066
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] tRNA遺伝子データベースの構築に向けた網羅的tRNA遺伝子の探索 (2007)
  • Author(s): 小原康雄、井口八郎、武藤あき, ら、山田優子、木ノ内誠、前野聖、金谷重彦、池村淑道、阿部貴志
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大会・合同年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-13
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Search for tRNA genes: construction of tRNA gene data-base (2007)
  • 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
• [Presentation] Trans-translation mediated by tmRNA and SmpB (2007)
  • 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
• [Presentation] Trans-translation mediated by tmRNA and SmpB (2007)
  • 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
• [Presentation] Interaction of SmpB with ribosome by directed hydroxyl radical Probing (2007)
  • Author(s): Kurita, D., Konno, T., Takada, K., Muto, A. & Himeno, H
  • Organizer: 22nd tRNA Workshop.
  • Place of Presentation: Uppsala, Sweden
  • Year and Date: 2007-11-05
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Interaction of SmpB with ribosome by directed Hyroxy radical probing (2007)
  • Author(s): Kimura, T., Konno, T., Takada, K., Muto, A. & Himeno, H
  • Organizer: The 22nd tRNA workshop
  • Place of Presentation: Uppsala, Sweden
  • Year and Date: 2007-11-05
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] tmRNA、SmpB、リボソーム間の相互作用の解析によるトランス・トランスレーションにおける翻訳再開機構の解明 (2007)
  • Author(s): 今野貴之、栗田大輔、武藤あき, ら、姫野俵太
  • Organizer: 第9回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 名古屋
  • Year and Date: 2007-07-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] リボソーム小サブユニット上におけるRsgAの相互作用部位の解析 (2007)
  • Author(s): 木村天胤、高木邦明、武藤あき, ら、姫野俵太
  • Organizer: 第9回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 名古屋
  • Year and Date: 2007-07-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] リボソーム小サブユニット依存GTPase RsgA欠損による浸透圧耐性の獲得 (2007)
  • Author(s): 長谷要一、武藤あき, ら、姫野俵太
  • Organizer: 第9回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 名古屋
  • Year and Date: 2007-07-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] リボソームタンパク質S1はトランス・トランスレーションの初期段階で重要でない (2007)
  • Author(s): 高田一馬、竹本千重、川添将仁、今野貴之、白水美香子、横山茂之、武藤あき, ら、姫野俵太
  • Organizer: 第9回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 名古屋
  • Year and Date: 2007-07-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] The mechnism of translatation resuming in trans-translation analyzed by interaction between tmRNA, SmpB and ribosome (2007)
  • Author(s): Konno, T., Kurita, D., 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
• [Presentation] RsgA binding site on 30S subunit of ribosome (2007)
  • Author(s): Kimura, T., Takagi, K., 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
• [Presentation] Deleting gene for RsgA, small subunit dependent GTPase, causes osmo-resistanc (2007)
  • Author(s): Hase, Y., 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
• [Presentation] Ribosomal rotein S1 is not involved in the early stage of trans-translation (2007)
  • 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
• [Presentation] Interaction between SmpB and ribosome from directed hydroxyl radical probing (2007)
  • Author(s): Muto, A., Kurita, D., Sasaki, R. & Himeno, H.
  • Organizer: Ribosomes 2007
  • Place of Presentation: Cape Cod, MA, USA
  • Year and Date: 2007-06-05
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Interaction between SmpB and ribosome from directed hydroxyl radical probing (2007)
  • Author(s): Muto, A., Kurita, D., Sasaki, R. & Himeno, H.
  • Organizer: Riboome 2007
  • Place of Presentation: Cape Cod, USA
  • Year and Date: 2007-06-05
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Expression of C. elegans novel ncRNAs. Regularoty RNAs (2006)
  • 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
• [Presentation] Expression of C. elegans novel ncRNAs: Regularoty RNAs (2006)
  • 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
• [Presentation] 高度好熱菌の無細胞翻訳系を用いたトランス・トランスレーション、(淡路)(2006). (2006)
  • Author(s): 高田一馬、竹本千重、州添将仁、今野貴之、浅野クリスナ、白水美香子、横山茂之、武藤あき, ら、姫野俵太
  • Organizer: 第8回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 淡路
  • Year and Date: 2006-07-18
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 新規抗生物質によるRsgAのGTP加水分解阻害メカニズムの解析 (2006)
  • Author(s): 木村天胤, 村上貴宣, 橋本 勝, 奥野智旦, 武藤あき, ら, 姫野俵太
  • Organizer: 第8回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 淡路
  • Year and Date: 2006-07-18
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Hydroxyradical fbotprintingによるSmpBとリボソームの相互作用の解明 (2006)
  • Author(s): 栗田大輔、今野貴之、佐々木瑠美、浅野クリスナ、高田一馬、武藤あき, ら、姫野俵太
  • Organizer: 第8回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 淡路
  • Year and Date: 2006-07-18
  • Description: 「研究成果報告書概要(和文)」より
• [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
• [Presentation] Inhibition of GTPase activity of RsgA by novel antibiotics (2006)
  • 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
• [Presentation] Interaction of SmpB with ribosome by hydroxyradical footprinting (2006)
  • 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
• [Presentation] 大腸菌リボソーム小サブユニットで活性化される新規GTPase(RsgA)に作用するCyclothiazomycin B1の作用 (2006)
  • Author(s): 木村天胤, 村上貴宣, 橋本 勝, 奥野智旦, 武藤あき, ら, 姫野俵太
  • Organizer: 日本農芸化学会2006年度大会
  • Place of Presentation: 京都
  • Year and Date: 2006-03-26
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Effect of Cyclothazomycin B1 on a novel GTPase activated by small ribosomal subunit, RsgA (2006)
  • 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
• [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
• [Presentation] Functional analysis of factors involved in trans-translation (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: Tokyo, Japan
  • Year and Date: 2005-10-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] ピューロマイシンによるリボソーム小サブユニット依存型GTPase欠損株の70S形成能の回復 (2005)
  • Author(s): 杉山和佳奈、武藤あき, ら、姫野俵太
  • Organizer: 第7回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 弘前
  • Year and Date: 2005-08-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] アミノグリコシド系抗生物質によるトランストランスレーション開始点のシフト (2005)
  • Author(s): 今野貴之、高橋千治、栗田大輔、浅野クリスナ、三浦比佳理、武藤あき, ら、姫野俵太
  • Organizer: 第7回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 弘前
  • Year and Date: 2005-08-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 翻訳がストールしたリボソーム中のA部位におけるtmRNA, tRNA, RF, RRFとの間の競合 (2005)
  • Author(s): 浅野クリスナ、今野貴之、栗田大輔、三浦比佳理、武藤あき, ら、姫野俵太
  • Organizer: 第7回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 弘前
  • Year and Date: 2005-08-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 枯草菌胞子形成過程におけるtmRNAによるトランス翻訳の関与 (2005)
  • Author(s): 阿部鉄平、榊和貴、藤原愛、大塚久志、氏家博美、牛田千里、姫野儀太、佐藤勉、武藤あき, ら
  • Organizer: 第7回RNAミーティング(日本RNA学会年会)
  • Place of Presentation: 弘前
  • Year and Date: 2005-08-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Puromycin restores defect of 70S ribosome formation caused by deficiency of 30S subunit dependent GTPase (2005)
  • Author(s): Sugiyama. W., 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
• [Presentation] The shift of trans-translation initiation by aminoglycosides (2005)
  • 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
• [Presentation] Competitions between tmRNA, tRNA, RF and RRF on the A-ste of stalled ribosome (2005)
  • 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
• [Presentation] Involvement of mRNA-mediated trans-translation in sporulation process of Bacillus subtilis (2005)
  • 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
• [Book] 細胞における蛋白質の一生-生成・成熟・輸送・管理・分解・病態-(翻訳と共役した蛋白質品質管理システム:トランスレーション)(小椋光、遠藤斗志也、森正敬、吉田賢右編)、 (2005)
  • Author(s): 姫野俵太、武藤あき, ら
  • Total Pages: 1062-1066
  • Publisher: 共立出版
  • Description: 「研究成果報告書概要(和文)」より