ペプチジルｔＲＮＡの脱落と新規翻訳制御機構の提唱

• Project Area: Diversity and asymmetry achieved by RNA program
• Project/Area Number: 23112705
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 鈴木 勉 東京大学, 工学(系)研究科(研究院), 教授 (20292782)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥18,889,000 (Direct Cost: ¥14,530,000、Indirect Cost: ¥4,359,000); Fiscal Year 2012: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2011: ¥9,529,000 (Direct Cost: ¥7,330,000、Indirect Cost: ¥2,199,000)
• Keywords: ペプチジルtRNA / Peptidyl-tRNA hydrolase / 翻訳校正機構 / リボソーム / peptidyl-tRNA hydrolase / RNAマススペクトロメトリー / 新生ペプチド鎖 / ペプチド鎖脱出トンネル / 品質管理

Outline of Annual Research Achievements

バクテリアにおいて、翻訳を開始したすべてのリボソームが終止コドンに到達するわけではなく、伸長過程の初期において新生ペプチド鎖が結合したペプチジルtRNA(pep-tRNA)がリボソームから脱離する現象(drop off)が知られている。脱離したペプチジルtRNAは細胞内ではPeptidyl-tRNA hydrolase(PTH)の作用によって代謝される。pthは必須遺伝子であり、PTHを失活させると短時間でpep-tRNAが蓄積し死に至ることから、pep-tRNAの脱離は偶発的なものではなく恒常的に生じている。我々は高感度質量分析法とRNA単離法を駆使することで、大腸菌pth温度感受性株におけるpep-tRNAの詳細なプロファイリングに成功した。実際に、N末にMetを持つジペプチドからペンタペプチド程度の短いペプチド鎖を有するpep-tRNAが観測された。また50Sのペプチド脱出トンネル、30Sの遺伝暗号解読中心それぞれを構成するrRNAに変異を導入したところ、pep-tRNAの脱離が誘発又は抑制される変異体を獲得できた。したがって翻訳初期段階において、pep-tRNAは新生ペプチド鎖とペプチド脱出トンネルとの相互作用、アンチコドン近傍と暗号解読中心との相互作用によってリボソーム上に留まっていると考えられる。また、新生ペプチド部分の配列解析からその多くはORFの開始近傍の配列に帰属できることが判明したが、大腸菌ORFの配列と一致しないペプチド鎖を有するpep-tRNAが検出された。詳細な解析から、これらのpep-tRNAはnon-cognateなアミノアシルtRNAがAサイトに入り、ペプチド転移反応によってPサイトtRNAのペプチド鎖を引き受けた後にdrop offしたものであることが判明した。この現象は、ペプチド転移反応以後における新しい翻訳校正機構であると考えている。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] A cyclic form of N^6-threonylcarbamoyladenosine as a widely distributed tRNA hypermodification (2013)
  • Author(s): Miyauchi, K., Kimura, S. and Suzuki, T.
  • Journal Title: Nature Chemical Biology Volume: 9 Issue: 2 Pages: 105-11
  • DOI: 10.1038/nchembio.1137
  • Peer Reviewed
• [Journal Article] Decoding system for the AUA codon by tRNAIle with the UAU anticodon in Myrcrplasma mobile (2013)
  • Author(s): Taniguchi T, Miyauchi K, Nakane D, Miyata M, Muto A, Nishimura S, Suzuki T
  • Journal Title: Nucleic Acids Research Volume: 41 Issue: 4 Pages: 2621-2631
  • DOI: 10.1093/nar/gks1344
  • Peer Reviewed
• [Journal Article] Decoding mechanism of non-universal genetic codes in Loligo bleekeri mitochondria (2013)
  • Author(s): Ohira T, Suzuki T (第2著者), 他5名
  • Journal Title: J . Biol. Chem Volume: 288 Issue: 11 Pages: 7645-7652
  • DOI: 10.1074/jbc.m112.439554
  • Peer Reviewed
• [Journal Article] Trmt61B is a methyltransferase responsible for 1-methyladenosine at position 58 of human mitochondrial tRNAs (2012)
  • Author(s): Chujo, T. and Suzuki, T.
  • Journal Title: RNA Volume: 22 Issue: 12 Pages: 2269-2276
  • DOI: 10.1261/rna.035600.112
  • Peer Reviewed
• [Journal Article] Structure-function analysis of human TYW2 enzyme required for the biosynthesis of a highly modified wybutosine (yW) base in phenylalanine-tRNA. (2012)
  • Author(s): Rodriguez, V., Vasudevan, S., Noma, A., Carlson, B.A., Green, J.E., Suzuki, T. and Chandrasekharappa, SC.
  • Journal Title: PLoS ONE Volume: 7 Issue: 6 Pages: e39297-e39297
  • DOI: 10.1371/journal.pone.0039297
  • Peer Reviewed
• [Journal Article] LRPPRC/SLIRP suppresses PNPase-mediated mRNA decay and promotes polyadenylation in human mitochondria. (2012)
  • Author(s): Chujo, T., Ohira, T., Sakaguchi, Y., Goshima, N., Nomura, N., Nagao, A. and Suzuki, T.
  • Journal Title: Nucleic Acids Res. Volume: 40 Issue: 16 Pages: 8033-8047
  • DOI: 10.1093/nar/gks506
  • Peer Reviewed
• [Journal Article] Base methylations in the double-stranded RNA by a fused methyltransferase bearing unwinding activity (2012)
  • Author(s): Kimura S., Ikeuchi Y., Kitahara K., Sakaguchi Y., Suzuki T., Suzuki T.
  • Journal Title: Nucleic Acids Research Volume: (掲載確定)(印刷中) Issue: 9 Pages: 4071-4085
  • DOI: 10.1093/nar/gkr1287
  • Peer Reviewed
• [Journal Article] Loss of ribosomal RNA modification causes developmental defects in zebrafish (2011)
  • Author(s): Higa-Nakamine S, Suzuki T (第2著者) 他7名
  • Journal Title: Nucleic Acids Research Volume: 40 Issue: 1 Pages: 391-398
  • DOI: 10.1093/nar/gkr700
  • Peer Reviewed
• [Journal Article] Molecular basis of dihydrouridine formation on tRNA (2011)
  • Author(s): Yu F, Tanaka Y, Yamashita K, Suzuki T (第4著者), 他4名
  • Journal Title: Proc Natl Acad Sci U S A Volume: 108 Issue: 49 Pages: 19593-19598
  • DOI: 10.1073/pnas.1112352108
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray diffraction analysis of an archaeal tRNA modification enzyme TiaS complexed with tRNA^<Ile2> and ATP (2011)
  • Author(s): Osawa, T., Inanaga, H., Kimura, S., Terasaka, N., Suzuki, T., Numata T.
  • Journal Title: Acta Crystallograph Sect F Struct Biol Cryst Commun Volume: 67 Issue: 11 Pages: 1414-1416
  • DOI: 10.1107/s1744309111034890
  • Peer Reviewed
• [Journal Article] Structural basis of tRNA agmatinylation essential for AUA codon decoding (2011)
  • Author(s): Osawa T., Kimura S., Terasaka N., Inanaga H., Suzuki T.
  • Journal Title: Nature Structural & Molcular Biology Volume: 18 Issue: 11 Pages: 1275-1280
  • DOI: 10.1038/nsmb.2144
  • Peer Reviewed
• [Journal Article] Biogenesis of 2-agmatinylcytidine catalyzed by the dual protein and RNA kinase TiaS (2011)
  • Author(s): Terasaka N., Kimura S., Osawa T., Numata T., Suzuki T.
  • Journal Title: Nature Structural & Molecular Biology Volume: 18 Issue: 11 Pages: 1268-1274
  • DOI: 10.1038/nsmb.2121
  • Peer Reviewed
• [Journal Article] Taurine-containing uridine modifications in tRNA anticodons are required to decipher non-universal genetic codes in ascidian mitochondria (2011)
  • Author(s): Suzuki T (第1著者), 他8名
  • Journal Title: J. Biol. Chem Volume: 286 Issue: 41 Pages: 35494-35498
  • DOI: 10.1074/jbc.m111.279810
  • Peer Reviewed
• [Journal Article] Human Mitochondrial tRNAs : Biogenesis, Function, Structural Aspects, and Diseases (2011)
  • Author(s): Suzuki T, Nagao A and Suzuki T
  • Journal Title: Annu. Rev. Genet Volume: 45 Issue: 1 Pages: 299-329
  • DOI: 10.1146/annurev-genet-110410-132531
  • Peer Reviewed
• [Journal Article] Deficit of tRNA(Lys) modification by Cdkall causes the development of type 2 diabetes in mice (2011)
  • Author(s): Wei FY., Suzuki T., Watanabe S., Kimura S., Kaitsuka T., Fujimura A., Matsui H., Atta M., Michiue H., Fontecave M., Yamagata K., Suzuki T., Tomizawa K.
  • Journal Title: The Journal of Clinical Investigation Volume: 121 Issue: 9 Pages: 3598-3608
  • DOI: 10.1172/jci58056
  • Peer Reviewed
• [Journal Article] Retrograde nuclear import of tRNA precursors is required for modified base biogenesis in yeast (2011)
  • Author(s): Ohira, T. and Suzuki, T.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 108 Issue: 26 Pages: 10502-10507
  • DOI: 10.1073/pnas.1105645108
  • Peer Reviewed
• [Journal Article] Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae (2011)
  • Author(s): Noma A, Yi S, Katoh T, Takai Y, Suzuki T and Suzuki T
  • Journal Title: RNA Volume: 17 Issue: 6 Pages: 1111-1119
  • DOI: 10.1261/rna.2653411
  • Peer Reviewed
• [Journal Article] Human mitochondrial diseases caused by lack of taurine-modification in mitochondrial tRNAs (2011)
  • Author(s): Suzuki T, Nagao A and Suzuki T
  • Journal Title: Wiley Interdiscip Rev RNA Volume: 2 Issue: 3 Pages: 376-386
  • DOI: 10.1002/wrna.65
  • Peer Reviewed
• [Presentation] RNAエピジェネティクスと生命現象 (2013)
  • Author(s): 鈴木 勉
  • Organizer: 日本化学会第93春季年会
  • Place of Presentation: 立命館大学びわこ・くさつキャンパス(滋賀)
  • Year and Date: 2013-03-22
  • Invited
• [Presentation] RNA修飾の多彩な機能と生命現象 (2013)
  • Author(s): 鈴木 勉
  • Organizer: 武田薬品湘南研究所セミナー
  • Place of Presentation: 武田薬品湘南研究所(藤沢)
  • Year and Date: 2013-03-13
  • Invited
• [Presentation] Molecular pathogenesis of human diseases associated with RNA modification disorder (2012)
  • Author(s): 鈴木 勉
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場・マリンメッセ福岡(福岡)
  • Year and Date: 2012-12-13
  • Invited
• [Presentation] A cyclic form of N6-threonylcarbamoyladenosine as a widely distributed tRNA hypermodification (2012)
  • Author(s): Tsutomu Suzuki
  • Organizer: XXIV tRNA Conference
  • Place of Presentation: Olmue, Chile
  • Year and Date: 2012-12-04
  • Invited
• [Presentation] Discovery and functional analysis of novel RNA modifications (2012)
  • Author(s): Tsutomu Suzuki
  • Organizer: Invited Seminar in Beijing Institute of Genetics
  • Place of Presentation: Beijing, China
  • Year and Date: 2012-10-15
  • Invited
• [Presentation] The cyclic form of N6-threonylcarbamoyladenosine (ct6A) as a bona fide hyper-modification of tRNAs in bacteria, lower eukaryotes and plants (2012)
  • Author(s): Tsutomu Suzuki
  • Organizer: Cold Spring Harbor Asia Conference RNA biology
  • Place of Presentation: Suzhou, China
  • Year and Date: 2012-10-10
  • Invited
• [Presentation] RNAとタンパク質をリン酸化する酵素の発見―RNA修飾の生合成と遺伝暗号の解読機構― (2012)
  • Author(s): 鈴木 勉
  • Organizer: 酵素工学研究会第68回講演会
  • Place of Presentation: 東大本郷キャンパス（東京）
  • Year and Date: 2012-10-05
  • Invited
• [Presentation] The cyclic form of N6-threonylcarbamoyladenosine (ct6A) as a bona fide hyper-modification of tRNAs in bacteria, lower eukaryotes and plants (2012)
  • Author(s): Tsutomu Suzuki
  • Organizer: FEBS International Workshop: New Developments in RNA Biology
  • Place of Presentation: Tavira, Portugal
  • Year and Date: 2012-09-02
  • Invited
• [Presentation] RNA modifications as naturally-selected chemical diversity involved in various biological processes (2011)
  • Author(s): Tsutomu Suzuki
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜(招待講演)
  • Year and Date: 2011-12-13
• [Presentation] ミトコンドリア脳筋症とtRNAのタウリン修飾欠損 (2011)
  • Author(s): 鈴木勉
  • Organizer: 第223回川崎医学会講演会
  • Place of Presentation: 倉敷(招待講演)
  • Year and Date: 2011-11-29
• [Presentation] RNA修飾の多彩な機能と生命現象 (2011)
  • Author(s): 鈴木勉
  • Organizer: 宮崎大学医学部講演会
  • Place of Presentation: 宮崎(招待講演)
  • Year and Date: 2011-11-24
• [Presentation] Biogenesis and function of 2-agmatinylcytidine found at the wobble position of archaeal tRNA^<Ile> (2011)
  • Author(s): Tsutomu Suzuki
  • Organizer: ISNAC2011第38回国際核酸化学シンポジウム
  • Place of Presentation: 札幌(招待講演)
  • Year and Date: 2011-11-11
• [Presentation] RNAマススペクトロメトリー (2011)
  • Author(s): 鈴木勉
  • Organizer: ヒューマンサイエンス財団セミナー
  • Place of Presentation: 東京(招待講演)
  • Year and Date: 2011-10-06
• [Presentation] RNA修飾の生合成から細胞内の化学反応を学ぶ (2011)
  • Author(s): 鈴木勉
  • Organizer: 生体機能関連化学部会「若手フォーラム」
  • Place of Presentation: つくば(招待講演)
  • Year and Date: 2011-09-11
• [Presentation] マイクロRNAと核酸医薬のマススペクトロメトリー (2011)
  • Author(s): 鈴木勉
  • Place of Presentation: 箱根(招待講演)
  • Year and Date: 2011-07-11
• [Remarks] タンパク質合成に必須なtRNAの修飾構造を解明 -40年前に決定された化学構造は分解物であった-
  • URL: http://www.t.u-tokyo.ac.jp/epage/release/2012/12122001.html
• [Remarks] 東大、タンパク質合成に必須なtRNAの新規修飾構造を発見・解明
  • URL: http://release.nikkei.co.jp/detail.cfm?relID=326829lindID=5
• [Remarks] タンパク質合成に必須なtRNA修飾の化学構造が決定された
  • URL: http://first.lifesciencedb.jp/archives/6418
• [Remarks] Essential tRNA modification stands in....
  • URL: http://59.106.161.11/en/todai-research/research-news/essential-trna-modification-stands-in-the-spotlight/
• [Remarks]
  • URL: http://www.u-tokyo.ac.jp/public/public01_231017_j.html
• [Remarks]
  • URL: http://first.lifesciencedb.jp/archives/3819#more-3819