Structural basis of G-quadruplex recognition by TLS protein which lead to telomere lengthening

• Project/Area Number: 18K14636
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Kyoto University
• Principal Investigator: Kondo Keiko 京都大学, エネルギー理工学研究所, 研究員 (00707474)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: NMR / 四重鎖核酸 / 核酸結合蛋白質 / 天然変性蛋白質

Outline of Final Research Achievements

Interaction of telomeric DNA and TERRA with peptide motifs in TLS, where tyrosine residue or phenylalanine residue is flanked with two RGG motifs, were analyzed by NMR. The peptide motif was suggested to bind to a G-tetrad and a loop region on the G-tetrad of telomeric DNA. On the other hand, it mainly bound to G-tetrads of TERRA. Additionally, the peptide motif was turned out to be able to interact with DNA-RNA hybrid quadruplex that formed by telomeric DNA and TERRA. Furthermore, conformational change of TLS from closed form to open form on binding telomeric DNA or TERRA was revealed.

Academic Significance and Societal Importance of the Research Achievements

TLSは細胞核内において遺伝子発現制御をはじめとする種々の制御に関与する核酸結合蛋白質であり、その機能不全は神経疾患をもたらすことも知られている。しかし、その核酸結合の機構は十分には明らかにされていない。本研究において得られたTLSと四重鎖核酸との結合に関する知見、および核酸との結合がTLSに構造変化をもたらすという知見は、生体内におけるTLSの機能を解明し、遺伝子発現制御などの普遍的な生命現象への理解を深めることに繋がる。

Research Products

• [Journal Article] RNA sequence and length contribute to RNA-induced conformational change of TLS/FUS (2020)
  • Author(s): Hamad Nesreen、Mashima Tsukasa、Yamaoki Yudai、Kondo Keiko、Yoneda Ryoma、Oyoshi Takanori、Kurokawa Riki、Nagata Takashi、Katahira Masato
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 2629-2629
  • DOI: 10.1038/s41598-020-59496-0
  • Peer Reviewed / Open Access
• [Journal Article] Versatility of RNA-Binding Proteins in Living Cells (2019)
  • Author(s): R. Kurokawa et al.
  • Journal Title: Biomedical Sciences Volume: 5 Issue: 1 Pages: 7-13
  • DOI: 10.11648/j.bs.20190501.12
  • Peer Reviewed / Open Access
• [Journal Article] NMR Analysis on Molecular Interaction of Lignin with Amino Acid Residues of Carbohydrate-Binding Module from Trichoderma reesei Cel7A (2019)
  • Author(s): Yuki Tokunaga, Takashi Nagata, Takashi Suetomi, Satoshi Oshiro, Keiko Kondo, Masato Katahira and Takashi Watanabe
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 1977-1977
  • DOI: 10.1038/s41598-018-38410-9
  • NAID: 120006554877
  • Peer Reviewed / Open Access
• [Journal Article] Identification of key residues for activities of atypical glutathione S-transferase of Ceriporiopsis subvermispora, a selective degrader of lignin in woody biomass, by crystallography and functional mutagenesis (2019)
  • Author(s): Osman Wan Hasnidah Wan、Mikami Bunzo、Saka Naoki、Kondo Keiko、Lin Meng-I、Nagata Takashi、Katahira Masato
  • Journal Title: International Journal of Biological Macromolecules Volume: 132 Pages: 222-229
  • DOI: 10.1016/j.ijbiomac.2019.03.199
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure of a serine-type glutathione S-transferase of Ceriporiopsis subvermispora and identification of the enzymatically important non-canonical residues by functional mutagenesis (2019)
  • Author(s): Wan Osman Wan Hasnidah、Mikami Bunzo、Saka Naoki、Kondo Keiko、Nagata Takashi、Katahira Masato
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 510 Issue: 1 Pages: 177-183
  • DOI: 10.1016/j.bbrc.2019.01.076
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Multiplicity in Long Noncoding RNA Biomedical Sciences (2018)
  • Author(s): 3.R. Kurokawa, R. Komiya, T. Oyoshi, Y. Matsuno, H. Tani, M. Katahira, K. Hitachi, Y. Iwashita, T. Yamashita, K. Kondo, R. Yoneda, Y. Yamaoki, N. Ueda, T. Mashima, N. Kobayashi, T. Nagata, A. Kiyoishi, M. Miyake, F. Kano, M. Murata1, N. Hamad, K. Sasaki, N. Shoji
  • Journal Title: Biomedical Sciences Volume: 4 Issue: 2 Pages: 18-23
  • DOI: 10.11648/j.bs.20180402.11
  • Peer Reviewed / Open Access
• [Presentation] Promotor associated non-coding RNA, pncRNA, can diminish the aggregation of TLS caused by shearing stress (2019)
  • Author(s): Hamad Nesreen、Mashima Tsukasa、Yamaoki Yudai、Kondo Keiko、Yoneda Ryoma、Oyoshi Takanori、Kurokawa Riki、Nagata Takashi、Katahira Masato
  • Organizer: 第42回分子生物学会
• [Presentation] NMR studies for the complex of TLS/FUS protein and G-quadruplexes of telomeric DNA and TERRA, which induces telomere shortening (2018)
  • Author(s): Keiko Kondo, Tsukasa Mashima, Takanori Oyoshi, Riki Kurokawa, Naohiro Kobayashi, Takashi Nagata, Masato Katahira
  • Organizer: 第２回長鎖非コードRNA勉強会
• [Presentation] Conformational change detection of translocated in liposarcoma, TLS, upon binding to promoter-associated non-coding RNA, pncRNA (2018)
  • Author(s): N. Hamad, T. Mashima, Y. Yamaoki, K. Kondo, H. Watanabe, T. Uchihashi, R. Kurokawa, T. Nagata, M. Katahira
  • Organizer: International Conference on Biochemistry, Molecular Biology and Biotechnology 2018
  • Int'l Joint Research