Systematic understanding of novel anti-virus system in a cell

• Project/Area Number: 25440013
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Shinkai Akeo 国立研究開発法人理化学研究所, 横山構造生物学研究室, 先任研究員 (10391989)
• Co-Investigator(Renkei-kenkyūsha): AGARI Yoshihiro 国立研究開発法人理化学研究所, 放射光科学総合研究センター, 特別研究員 (50469889)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: CRISPR-Cas / 獲得免疫 / 超分子複合体 / 電子顕微鏡構造 / タンパク質 / 低分子RNA / リボヌクレアーゼ / Thermus thermophilus / 国際情報交換（オランダ、アメリカ） / CRISPR-Casシステム

Outline of Final Research Achievements

CRISPR-Cas system is a bacterial adaptive immune system, which degrades foreign DNA or RNA such as that derived from virus. We selected a thermophilic bacterium, Thermus thermophilus, which has several CRISPR-Cas systems, for a model organism to study their systematic manner in a cell. We investigated structure and function of two large complexes comprising two CRISPR-Cas systems of the strain. We found that each complex was composed of several protein subunits and a small RNA molecule, and adopted a helical structure. Nucleotide sequences of the small RNA molecules are similar to each other. Both complexes degraded single-strand RNA in a similar manner. These results suggest that the two complexes have been evolved from a common ancestor, and that they comprise backup system in a cell to respond to virus infection.

Research Products

• [Int'l Joint Research] Wageningen University/University of Utrecht(オランダ)
• [Int'l Joint Research] University of California, Berkeley(米国)
• [Journal Article] Structure and function of Type III-B CRISPR-Cmr complex from Thermus thermophilus (2015)
  • Author(s): 新海 暁男
  • Journal Title: 生化学 Volume: 87 Issue: 4 Pages: 454-458
  • DOI: 10.14952/SEIKAGAKU.2015.870454
  • ISSN: 0037-1017
  • Year and Date: 2015-08-25
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structures of the CRISPR-Cmr complex reveal mode of RNA target positioning. (2015)
  • Author(s): Taylor D.W., Zhu Y., Staals R.H.J., Kornfeld J.E., Shinkai A., van der Oost J., Nogales E., Doudna J.A.
  • Journal Title: Science Volume: 348 Issue: 6234 Pages: 581-585
  • DOI: 10.1126/science.aaa4535
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structure and Function of CRISPR-Cas System (2014)
  • Author(s): 新海 暁男
  • Journal Title: Seibutsu Butsuri Volume: 54 Issue: 5 Pages: 247-252
  • DOI: 10.2142/biophys.54.247
  • NAID: 130004693950
  • ISSN: 0582-4052, 1347-4219
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] RNA-targeting by the Type III-A CRISPR-Cas complex of Thermus thermophilus. (2014)
  • Author(s): Staals R.H.J., Zhu Y., Taylor D.W., Kornfeld J.E., Sharma K., Barendregt A., Koehorst J.J., Vlot M., Varossieau K., Neupane N., Sakamoto K., Suzuki T., Dohmae N., Yokoyama S., Schaap P.J., Urlaub H., Heck A.J.R., Nogales E., Doudna J.A., Shinkai A., van der Oost J.
  • Journal Title: Molecular Cell Volume: 56 Issue: 4 Pages: 518-530
  • DOI: 10.1016/j.molcel.2014.10.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus (2013)
  • Author(s): Staals, R. H. J., Agari, Y., Maki-Yonekura, S., Zhu, Y., Taylor, D. W., van Duijn, E., Barendregt, A., Vlot, M., Koehorst, J. J., Sakamoto, K., Masuda, A., Dohmae, N., Schaap, P. J., Doudna, J. A., Heck, A. J. R., Yonekura, K., van der Oost, J. and Shinkai, A.
  • Journal Title: Molecular Cell Volume: 52 Issue: 1 Pages: 135-145
  • DOI: 10.1016/j.molcel.2013.09.013
  • Peer Reviewed
• [Presentation] 高度好熱菌Thermus thermophilusのType III CRISPR-Casシステムを構成しているリボヌクレオタンパク質複合体の構造と機能 (2015)
  • Author(s): 新海 暁男
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド、兵庫県神戸市
  • Year and Date: 2015-12-03
• [Presentation] 高度好熱菌Thermus thermophilus由来Type III-B CRISPR-Casシステムを構成するCmr複合体の構造と機能 (2015)
  • Author(s): 新海 暁男
  • Organizer: 第17回日本RNA学会年会
  • Place of Presentation: ホテルライフォート札幌、北海道札幌市
  • Year and Date: 2015-07-15
• [Presentation] The Cmr complex of Thermus thermophilus
  • Author(s): Raymond H.J. Staals, Yoshihiro Agari, John van der Oost, Akeo Shinkai
  • Organizer: CRISPR: evolution, mechanisms and infection
  • Place of Presentation: University of St Andrews, United Kingdom
• [Presentation] Functional and structural genomics studies on Thermus thermophilus CRISPR-Cas systems
  • Author(s): Akeo Shinkai
  • Organizer: The 65th Fujihara Seminar, Internal Symposium on Synthetic Biology of Unnatural Base Pairs and Amino Acids, The Fujihara Foundation of Science
  • Place of Presentation: グランドホテルニュー王子、北海道苫小牧市
• [Remarks] 理化学研究所 プレスリリース：獲得免疫の起源を探る
  • URL: http://www.riken.jp/pr/press/2013/20131011_1/
• [Remarks] 理化学研究所 60秒でわかるプレスリリース：獲得免疫の起源を探る
  • URL: http://www.riken.jp/pr/press/2013/20131011_1/digest/