Transcription-coupled pre-mRNA processing in living animal

• Project/Area Number: 17H03633
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: KUROYANAGI Hidehito 東京医科歯科大学, 難治疾患研究所, 准教授 (30323702)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2018: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2017: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 遺伝子発現制御 / 新生RNA / 線虫 / スプライシング / 転写後プロセシング / mRNA / 代謝標識 / RNA-seq解析 / RNA-seq / mRNA前駆体 / RNAプロセシング / 転写 / 組織特異性 / RNAseq / 遺伝子発現

Outline of Final Research Achievements

We developed new methods to label and purify nascent precursor messenger RNAs (pre-mRNAs) in a nematode worm C. elegans. With the methods, we revealed dynamics of pre-mRNA processing such as trans-splicing, cis-splicing and cleavage at a polyadenylation site for the first time in multicellular organisms. We also analyzed nascent pre-mRNAs in a splicing factor mutant and revealed effects of the factor on the dynamics of pre-mRNA processing.

Academic Significance and Societal Importance of the Research Achievements

mRNA前駆体のプロセシング動態が多細胞生物の個体レベルで明らかになるのは初めてである。哺乳類ではイントロンが長大であることから、同様の方法を適用しても同様の結果を得ることは実質的に困難である。本研究成果は、哺乳類を含む他の多細胞動物における転写と共役したmRNAの転写後プロセシングによる遺伝子発現制御機構を考察する上で基礎となる重要な知見を与えるものである。

Research Products

• [Int'l Joint Research] Emory University(米国)
• [Int'l Joint Research] University of Pittsburgh(米国)
• [Int'l Joint Research] University of California at Los Angeles/Emory University/University of Pittsburgh(米国)
• [Int'l Joint Research] Institut Europien de Chimie & Biologie(フランス)
• [Int'l Joint Research] Emory University(米国)
• [Int'l Joint Research] University of California, Los Angeles(米国)
• [Int'l Joint Research] Indiana University School of Medicine(米国)
• [Journal Article] mRNA Editing, Processing, and Quality Control in Caenorhabditis elegans (2020)
  • Author(s): Joshua A. Arribere, Hidehito Kuroyanagi, and Heather A. Hundley
  • Journal Title: Genetics Volume: 215 Issue: 3 Pages: 1-39
  • DOI: 10.1534/genetics.119.301807
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Alternative splicing of the Caenorhabditis elegans lev-11 tropomyosin gene is regulated in a tissue-specific manner (2018)
  • Author(s): Watabe Eichi、Ono Shoichiro、Kuroyanagi Hidehito
  • Journal Title: Cytoskeleton Volume: 75 Issue: 10 Pages: 427-436
  • DOI: 10.1002/cm.21489
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Phosphorylation of the RSRSP stretch is critical for splicing regulation by RNA-Binding Motif Protein 20 (RBM20) through nuclear localization (2018)
  • Author(s): Rie Murayama, Mariko Kimura-Asami, Marina Togo-Ohno, Yumiko Yamasaki-Kato, Taeko K. Naruse, Takeshi Yamamoto, Takeharu Hayashi, Tomohiko Ai, Katherine G. Spoonamore, Richard J. Kovacs, Matteo Vatta, Mai Iizuka, Masumi Saito, Shotaro Wani, Yuichi Hiraoka, Akinori Kimura & Hidehito Kuroyanagi
  • Journal Title: Scientific Reports Volume: 印刷中 Issue: 1 Pages: 8970-8983
  • DOI: 10.1038/s41598-018-26624-w
  • NAID: 120007031665
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tropomyosin isoforms differentially affect muscle contractility in the head and body regions of the nematode Caenorhabditis elegans (2018)
  • Author(s): Barnes Dawn E.、Watabe Eichi、Ono Kanako、Kwak Euiyoung、Kuroyanagi Hidehito、Ono Shoichiro
  • Journal Title: Molecular Biology of the Cell Volume: 29 Issue: 9 Pages: 1075-1088
  • DOI: 10.1091/mbc.e17-03-0152
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Alternative Splicing Regulator RBM20 and Cardiomyopathy (2018)
  • Author(s): Watanabe Takeshi、Kimura Akinori、Kuroyanagi Hidehito
  • Journal Title: Frontiers in Molecular Biosciences Volume: 5 Pages: 105-105
  • DOI: 10.3389/fmolb.2018.00105
  • NAID: 120007150215
  • Peer Reviewed / Open Access
• [Journal Article] Phosphorylation of the RSRSP stretch is critical for splicing regulation by RNA-Binding Motif Protein 20 (RBM20) through nuclear localization (2018)
  • Author(s): Rie Murayama, Mariko Kimura-Asami, Marina Togo-Ohno, Yumiko Yamasaki-Kato, Taeko K. Naruse, Takeshi Yamamoto, Takeharu Hayashi, Tomohiko Ai, Katherine G. Spoonamore, Richard J. Kovacs, Matteo Vatta, Mai Iizuka, Masumi Saito, Shotaro Wani, Yuichi Hiraoka, Akinori Kimura and Hidehito Kuroyanagi
  • Journal Title: Scientific Reports Volume: 印刷中
  • NAID: 120007031665
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] An emerging model organism Caenorhabditis elegans for alternative pre-mRNA processing in vivo (2017)
  • Author(s): Shotaro Wani & Hidehito Kuroyanagi
  • Journal Title: Wiley Interdisciplinary Reviews: RNA Volume: 印刷中 Issue: 6
  • DOI: 10.1002/wrna.1428
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Genome-Wide Kinetic Analysis of pre-mRNA Processing in C. elegans (2019)
  • Author(s): Eichi Watabe and Hidehito Kuroyanagi
  • Organizer: The 24th Annual Meeting of the RNA Society
  • Int'l Joint Research
• [Presentation] Genome-Wide Kinetic Analysis of pre-mRNA Processing in C. elegans (2019)
  • Author(s): Eichi Watabe and Hidehito Kuroyanagi
  • Organizer: The 22nd International C. elegans Meeting
  • Int'l Joint Research
• [Presentation] PES-4 regulates head-muscle-specific alternative splicing of the tropomyosin pre-mRNA (2019)
  • Author(s): Eichi Watabe and Hidehito Kuroyanagi
  • Organizer: The 22nd International C. elegans Meeting
  • Int'l Joint Research
• [Presentation] Homeostasis of S-adenosylmethionine (SAM) synthetase genes in C. elegans by alternative pre-mRNA splicing through m6A modification at the 3' spice site (2019)
  • Author(s): Shotaro Wani, Eichi Watabe, Yuma Ishigami, Marina Togo-Ohno, Tsutomu Suzuki and Hidehito Kuroyanagi
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] Genome-Wide Kinetic Analysis of pre-mRNA Processing Reveals Action of Splicing Regulators (2019)
  • Author(s): Hidehito Kuroyanagi
  • Organizer: Polish-Japanese RNA Meeting
  • Int'l Joint Research / Invited
• [Presentation] トロポミオシンをコードするlev-11遺伝子の選択的スプライシング制御 (2019)
  • Author(s): 黒柳秀人
  • Organizer: 東京地区線虫勉強会
• [Presentation] 動物個体におけるmRNA前駆体プロセシング動態の解析 (2018)
  • Author(s): 渡部栄地，黒柳秀人
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] Genome-wide Kinetic Analysis of Pre-mRNA Processing in Living Animals (2018)
  • Author(s): 渡部栄地，黒柳秀人
  • Organizer: 線虫研究の未来を創る会
• [Presentation] C. elegans代謝関連遺伝子のスプライシング制御における食餌の影響 (2018)
  • Author(s): 和仁翔太郎
  • Organizer: 東京地区線虫勉強会
• [Presentation] Genome-Wide Kinetic Analysis of Pre-mRNA Processing in Living Animals (2018)
  • Author(s): 渡部栄地
  • Organizer: 東京地区線虫勉強会
• [Presentation] 個体レベルで行うmRNA前駆体プロセシングの動態の解析 (2017)
  • Author(s): 渡部栄地、黒柳秀人
  • Organizer: 第19回日本RNA学会年会
• [Presentation] C. elegans代謝関連遺伝子のスプライシング制御における食餌の影響 (2017)
  • Author(s): 和仁翔太郎、黒柳秀人
  • Organizer: 第19回日本RNA学会年会
• [Presentation] Genome-wide kinetic analysis of pre-mRNA processing in living animals (2017)
  • Author(s): Eichi Watabe, Hidehito Kuroyanagi
  • Organizer: CSHL Meeting on 40 YEARS OF mRNA SPLICING: FROM DISCOVERY TO THERAPEUTICS
• [Presentation] 個体レベルで行う mRNA前駆体プロセシングの動態の解析 (2017)
  • Author(s): 渡部栄地、黒柳秀人
  • Organizer: ConBio2017（2017年度生命科学系学会合同年次大会）
• [Book] 遺伝子発現制御機構 (2017)
  • Author(s): 田村隆明、浦聖惠（編）
  • Total Pages: 264
  • Publisher: 東京化学同人
  • ISBN: 9784807909179
• [Remarks] 東京医科歯科大学 難治疾患研究所 フロンティア研究室 遺伝子発現制御学
• [Remarks] 東京医科歯科大学難治疾患研究所フロンティア研究室遺伝子発現制御学
• [Remarks] フロンティア研究室 遺伝子発現制御学
  • URL: http://www.tmd.ac.jp/end/
• [Remarks] 「ノンコーディングRNAネオタクソノミ」研究成果紹介
  • URL: https://ncrna.jp/blog/item/370-rouge
• [Remarks] みらいぶプラス
  • URL: https://www.milive-plus.net/newleader/049/