出芽酵母の胞子形成時におけるRNAエピジェネティクス

2014 Fiscal Year Annual Research Report

• Project/Area Number: 25620125
• 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): 2013-04-01 – 2015-03-31
• Keywords: RNA修飾 / m6A / 胞子形成

Outline of Annual Research Achievements

本研究では、出芽酵母の胞子形成時にのみ出現するmRNA上のm6A修飾の役割を明らかにすることを目的とし、m6A部位の探索を行う。胞子形成初期段階に特異的に増加するm6A修飾部位を網羅的に特定するために、抗m6A抗体を用いた免疫沈降法と次世代シークエンス解析を組み合わせたm6A-seqによる解析を行っている。実際に、feasibility studyにおいて、3,000遺伝子中に8,000か所以上のm6A候補領域を特定することに成功した。しかし、cDNAライブラリ調製の過程で転写産物の方向の情報が失われることや、解析した平均cDNA長が300塩基と長く検出したピークがブロードでありため、修飾部位の特定には至らなかった。より精度の高いデータを得るためには、RNAの断片化、免疫沈降、Strand-specificなcDNAライブラリの調製など、各ステップについて条件検討を行い最適化する必要がある。また本研究の過程において、市販されている抗m6A抗体の特異性が低いことが明らかとなった。今後は、より特異性の高い抗m6Aモノクローナル抗体の取得をめざし、m6A-seqを最適化する予定である。

Research Products

• [Journal Article] Iron-sulfur proteins responsible for RNA modifications (2015)
  • Author(s): Kimura, S. and Suzuki, T.
  • Journal Title: Biochimica et Biophysica Acta (BBA)-Molecular Cell Research Volume: 1853 Pages: 1272-1283
  • DOI: 10.1016/j.bbamcr.2014.12.010
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Rectifier of aberrant mRNA splicing recovers tRNA modification in familial dysautonomia (2015)
  • Author(s): Yoshida, M., Kataoka, N., Miyauchi, K., Ohe, K., Iida, K., Yoshida, S., Nojima, T., Okuno, Y., Onogi, H., Usui, T., Takeuchi, A., Hosoya, T., Suzuki, T. and Hagiwara, M.
  • Journal Title: Proc. Natl. Acad. Sci. USA. Volume: 112 Pages: 2764-2769
  • DOI: 10.1073/pnas.1415525112
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cdk5rap1-mediated 2-methylthio modification of mitochondrial tRNAs governs precise mitochondrial protein translation and contributes to myopathy in mice and humans (2015)
  • Author(s): Wei, F., Zhou, B., Suzuki, T., Miyata, K., Ujihara, Y., Takahashi, N., Xie, P., Michiue, H., Fujimura, A., Kaitsuka, T., Matsui, H., Koga, Y., Mohri, S., Suzuki, T., Oike, Y. and Tomizawa, K.
  • Journal Title: Cell Metabolism Volume: 21 Pages: 428-442
  • DOI: 10.1016/j.cmet.2015.01.019.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Defining fundamental steps in the assembly of Drosophila RNAi enzyme complex (2015)
  • Author(s): Iwasaki, S., Sasaki, H. M., Sakaguchi, Y., Suzuki, T., Tadakuma, H. and Tomari, Y.
  • Journal Title: Nature Volume: in press Pages: in press
  • DOI: 10.1038/nature14254
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A complete landscape of post-transcriptional modifications in mammalian mitochondrial tRNAs (2014)
  • Author(s): Suzuki, T. and Suzuki, T.
  • Journal Title: Nucleic Acids Res Volume: 42 Pages: 7346-7357
  • DOI: 10.1093/nar/gku390
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Discovery of the β-barrel-type RNA methyltransferase responsible for N6-methylation of N6-threonylcarbamoyladenosine in tRNAs (2014)
  • Author(s): Kimura, S., Miyauchi, K., Ikeuchi, Y., Thiaville, P.C., de Crecy-Lagard, and Suzuki, T.
  • Journal Title: Nucleic Acids Res Volume: 42 Pages: 9350-9365
  • DOI: 10.1093/nar/gku618
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A single acetylation of 18S rRNA is essential for biogenesis of the small ribosomal subunit in Saccharomyces cerevisiae (2014)
  • Author(s): Ito, S., Akamatsu, Y., Noma, A., Kimura, S., Miyauchi, K., Ikeuchi, Y., Suzuki, T. and Suzuki, T
  • Journal Title: J. Biol. Chem. Volume: 289 Pages: 26201-26212
  • DOI: 10.1074/jbc.M114.593996
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Human NAT10 is an ATP-dependent RNA acetyltransferase responsible for N4-acetylcytidine formation in 18S rRNA (2014)
  • Author(s): Ito, S., Horikawa, S., Suzuki, T., Kawauchi, H., Tanaka, Y., Suzuki, T. and Suzuki, T.
  • Journal Title: J. Biol. Chem. Volume: 289 Pages: 35724-35730
  • DOI: 10.1074/jbc.C114.602698
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] 5-Hydroxymethylcytosine plays a critical role in glioblastomagenesis by recruiting the CHTOP-methylosome Complex (2014)
  • Author(s): Takai, H., Masuda, K., Sato, T., Sakaguchi, Y., Suzuki, T., Suzuki, T., Koyama-Nasu, R., Nasu-Nishimura, Y., Katou, Y., Ogawa, H., Morishita, Y., Kozuka-Hata, H., Oyama, M., Todo, T., Ino, Y., Mukasa, A., Saito, N., Toyoshima, C., Shirahige, K. and Akiyama, T.
  • Journal Title: Cell Reports Volume: 9 Pages: 48-60
  • DOI: 10.1016/j.celrep.2014.08.071
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Convergent evolution of AUA decoding in bacteria and archaea (2014)
  • Author(s): Suzuki, T. and Numata, T.
  • Journal Title: RNA Biology Volume: 11 Pages: 1586-1596
  • DOI: 10.4161/15476286.2014.992281
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Essential RNA acetyltransferase required for ribosome biogenesis and tRNA modification (2015)
  • Author(s): Tsutomu Suzuki
  • Organizer: GRC on RNA editing and modification
  • Place of Presentation: Lucca, Italy
  • Year and Date: 2015-03-09
  • Invited
• [Presentation] RNA修飾の多彩な機能と疾患 (2015)
  • Author(s): 鈴木 勉
  • Organizer: 埼玉医科大学ゲノム医学研究センター学術集会
  • Place of Presentation: 埼玉医科大学ゲノム医学研究センター、埼玉県日高市
  • Year and Date: 2015-01-27
  • Invited
• [Presentation] Mitochondrial diseases caused by deficient tRNA modifications (2014)
  • Author(s): Tsutomu Suzuki
  • Organizer: AussieMit2014
  • Place of Presentation: Perth, Australia
  • Year and Date: 2014-12-01
  • Invited
• [Presentation] A single acetylation of 18S rRNA regulates ribosome biogenesis by sensing cellular energy status (2014)
  • Author(s): Tsutomu Suzuki
  • Organizer: Joint Australia and Japan RNA meeting (JAJRNA2014)
  • Place of Presentation: Sydney, Australia
  • Year and Date: 2014-11-05
  • Invited
• [Presentation] Biogenesis and function of cyclic t6A and derivatives (2014)
  • Author(s): Tsutomu Suzuki
  • Organizer: 25th tRNA conference
  • Place of Presentation: Kyllini, Greece
  • Year and Date: 2014-09-22
  • Invited
• [Presentation] ヒトはなぜ病気になるのでしょう？～病気の原因を分子レベルで探る～ (2014)
  • Author(s): 鈴木 勉
  • Organizer: 高校生のためのオープンキャンパス2014
  • Place of Presentation: 東京大学本郷キャンパス、東京
  • Year and Date: 2014-08-06
  • Invited
• [Book] RNAの修飾欠損と疾患/ファルマシア (2015)
  • Author(s): 鈴木 勉、浅野 奏、鈴木健夫
  • Total Pages: 5
  • Publisher: 公益社団法人 日本薬学会