Elucidation of inhibitory mechanism of binding of long noncoding RNA via arginine methylation and its physiological significance

• Project/Area Number: 18K06939
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 48040:Medical biochemistry-related
• Research Institution: Saitama Medical University
• Principal Investigator: KUROKAWA Riki 埼玉医科大学, 医学部, 教授 (70170107)
• Project Period (FY): 2018-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: アルギニンメチル化 / TLS / FUS / lncRNA / pncRNA-D / 相分離 / HAT / PRMT1 / RNA結合タンパク質 / RNA結合たんぱく質 / arginine / methylation / long noncoding RNA / histone acetylation

Outline of Final Research Achievements

Our previous data demonstrate that RNA-binding protein (RBP) TLS/FUS binds promoter-associated noncoding RNA-D (pncRNA-D) and strongly represses the CBP/p300 histone acetyltransferase (HAT) activity (Wang et al. Nature 2008). Recently, we have shown that protein arginine (R) methyltransferase 1(PRMT1)methylates arginine residues of the C-terminus of TLS and this R methylation represses RNA binding of TLS and also its inhibitory effect on the HAT activity. These data present a novel and comprehensive model showing that R methylation of TLS modulates its function and also regulates expression of relevant genes in the human genome.

Academic Significance and Societal Importance of the Research Achievements

これまで、TLS/FUSを含む多くのRNA結合タンパク質(RBP)が高度にアルギニン(R)メチル化されていることが知られていたが、その生理的な意義は大部分不明であった。今回の成果で、TLSのRメチル化が、RBPの中心的な機能であるRNA結合を制御することが明らかになった学術的な意義は大きい。今後は、この現象がほかのRBPでも機能していることをゲノムワイドで解明していきたい。一方、タンパク質のRメチル化は、大腸がんの発症や、胎生期の大脳の発達、iPS細胞の万能性の維持などに必須であり、これらの分子機構を解明することは社会的な要求に貢献することになろう。

Research Products

• [Journal Article] Identification of Long Noncoding RNA Recognized by RNA-Binding Protein TLS/FUS: Purification of RNAs by Affinity Chromatography of GST-TLS (2022)
  • Author(s): Naomi Ueda,Ryoma Yoneda,Riki Kurokawa
  • Journal Title: Biomedical Sciences Volume: 8 Pages: 144-156
  • Peer Reviewed / Open Access
• [Journal Article] m 6 A Modified Short RNA Fragments Inhibit Cytoplasmic TLS/FUS Aggregation Induced by Hyperosmotic Stress (2021)
  • Author(s): Ryoma Yoneda, Naomi Ueda, Riki Kurokawa
  • Journal Title: International Journal of Molecular Sciences Volume: Vol.22 Issue: 20 Pages: 11014-11014
  • DOI: 10.3390/ijms222011014
  • Peer Reviewed / Open Access
• [Journal Article] Non-coding RNA suppresses FUS aggregation caused by mechanistic shear stress on pipetting in a sequence-dependent manner (2021)
  • Author(s): Hamad Nesreen、Yoneda Ryoma、So Masatomo、Kurokawa Riki、Nagata Takashi、Katahira Masato
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 9523-9523
  • DOI: 10.1038/s41598-021-89075-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Potential Inhibitor Against Phase Separation, 1,6-hexanediol Specifically Binds to Beta Actin in Nuclear Extract of Human Cell Line (2020)
  • Author(s): Naomi Ueda, Yuki Hirose, Ryoma Yoneda, Toshikazu Bando, Riki Kurokawa
  • Journal Title: Biomedical Sciences Volume: 6 Pages: 88-97
  • Peer Reviewed / Open Access
• [Journal Article] Long noncoding RNA pncRNA-D?reduces cyclin D1 gene expression and arrests cell cycle through RNA m6A modification (2020)
  • Author(s): Yoneda Ryoma、Ueda Naomi、Uranishi Kousuke、Hirasaki Masataka、Kurokawa Riki
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 17 Pages: 5626-5639
  • DOI: 10.1074/jbc.ra119.011556
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Arginine methylation of translocated in liposarcoma (TLS) inhibits its binding to long noncoding RNA, abrogating TLS-mediated repression of CBP/p300 activity (2018)
  • Author(s): Cui Wei、Yoneda Ryoma、Ueda Naomi、Kurokawa Riki
  • Journal Title: Journal of Biological Chemistry Volume: 293 Issue: 28 Pages: 10937-10948
  • DOI: 10.1074/jbc.ra117.000598
  • Peer Reviewed
• [Journal Article] Affinity Profiles Categorize RNA-Binding Proteins into Distinctive Groups. (2018)
  • Author(s): Naomi Ueda、Riki Kurokawa
  • Journal Title: Biomedical Sciences Volume: 4 Pages: 24-31
  • Peer Reviewed / Open Access
• [Presentation] Quest for novel long non-coding RNAs with inhibitory activity against phase separation (2022)
  • Author(s): 黒川理樹
  • Organizer: 第45回分子生物学会年会
  • Invited
• [Presentation] Transcription regulatory lncRNA represses phase separation of TLS/FUS-Molecular linkage of lncRNA to phase separation by TLS in cellular functions (2021)
  • Author(s): 黒川 理樹
  • Organizer: 第44回日本分子生物学会年会
  • Invited
• [Presentation] 相分離・凝集体形成の抑制因子1,6-hexanediolの作用機構と生理的意義の解明 (2020)
  • Author(s): 黒川 理樹
  • Organizer: 第43回日本分子生物学会年会
  • Int'l Joint Research
• [Presentation] 多機能分子FUS/TLS-脂肪肉腫の融合遺伝子、家族性ALSの原因遺伝子、そして、相分離の中心分子としての役割 (2019)
  • Author(s): 黒川 理樹
  • Organizer: 第42回日本分子生物学会年会
  • Invited
• [Presentation] Arginine methylation of RNA binding protein TLS inhibits binding to long noncoding RNA, repressing the histone acetyltransferase activity. (2019)
  • Author(s): 黒川 理樹
  • Organizer: Keystone Symposia
  • Int'l Joint Research / Invited
• [Presentation] Arginine methylation of RNA binding protein TLS inhibits binding to long noncoding RNA, repressing the histone acetyltransferase activity アルギニンメチル化によるRNA結合タンパク質TLS機能の制御機構 (2018)
  • Author(s): 黒川 理樹
  • Organizer: 第41回日本分子生物学会年会
  • Invited