A novel RNA metabolism involved in cell differentiation and survival

• Project/Area Number: 25460371
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kochi University
• Principal Investigator: Shuji Sakamoto 高知大学, 教育研究部医療学系基礎医学部門, 准教授 (80397546)
• Research Collaborator: Todaka Hiroshi 高知大学, 教育研究部医療学系基礎医学部門, 助教 (80769662) ; Higuchi Takuma 高知大学, 教育研究部医療学系基礎医学部門, 助教 (10754567)
• 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: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 二本鎖RNA結合蛋白質 / microRNA生合成 / miRNA / 筋分化 / 中心核病 / Dynamin 2 / NF90 / NF45 / 筋再生 / 中心核 / 骨格筋 / 筋成熟 / マイクロRNA / Dynamin / 筋萎縮 / ミオパチー / Dynamin2

Outline of Final Research Achievements

1. The nuclear factor 90 (NF90) or the complex of NF90 with NF45 is known to be involved in microRNA biogenesis, mRNA stabilization, translational repression, replication of viral RNA and transcription in vitro. In this study, we found that the NF90-NF45 complex induces centronuclear myopathy through increased expression of dynamin 2, a causative gene of the disease, by an NF90-NF45-induced reduction of miR-133a expression in vivo.

2. We observed that the NF90-overexpressing HEK293 cells did not exhibit longer-term survival than the control cells transfected with mock plasmids when glucose was withdrawn, suggesting that cellular energy balance was not influenced by overexpression of NF90 in cultured cells in vitro.

Research Products

• [Journal Article] PNPLA3 is involved in hepatic fatty acid and triglyceride metabolism through XBP1 and modulation of endoplasmic reticulum stress in mice. (2016)
  • Author(s): Ochi T, Munekage K, Ono M, Higuchi T, Tsuda M, Hayashi Y, Okamoto N, Toda K, Sakamoto S, Oben JA, Saibara T
  • Journal Title: Hepatol Res Volume: 印刷中 Issue: 6 Pages: 584-592
  • DOI: 10.1111/hepr.12587
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Expression and phosphorylation state analysis of intracellular protein kinases using Multi- PK antibody and Phos- tag SDS- PAGE. (2015)
  • Author(s): Sugiyama, Y., Katayama, S., Kameshita, I., Morisawa, K., Higuchi, T., Todaka, H., Eiji Kinoshita, Kinoshita- Kikuta, E., Koike, T., Taniguchi, T., Sakamoto, S.
  • Journal Title: MethodsX Volume: 2 Pages: 469-474
  • DOI: 10.1016/j.mex.2015.11.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] New method for analysis of intracellular phosphorylation dynamics using Multi-PK antibody and Phos-tag (2015)
  • Author(s): 1. 杉山 康憲、亀下 勇、坂本 修士
  • Journal Title: Electrophoresis Letters Volume: 59 Issue: 2 Pages: 82-84
  • DOI: 10.2198/electroph.59.82
  • NAID: 130005103931
  • ISSN: 2189-2628, 2189-2636
  • Open Access
• [Journal Article] Overexpression of NF90-NF45 complex functions as a negative regulation in the microRNA precessing pathway. (2015)
  • Author(s): Todaka H, Higuchi T, Yagyu KI, Sugiyama Y, Yamaguchi F, Morisawa K, Ono M, Fukushima A, Tsuda M, Taniguchi T, Sakamoto S.
  • Journal Title: Molecular and Cellular Biology Volume: 印刷中
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Bofutsushosan, a Japanese herbal (Kampo) medicine, attenuates progression of nonalcoholic steatohepatitis in mice (2013)
  • Author(s): Masafumi Ono, Mitsunari Ogasawara, Akira Hirose, Sachiko Mogami, Nobuhiro Ootake, Kosuke Aritake, Takuma Higuchi, Nobuto Okamoto, Shuji Sakamoto, Masahiro Yamamoto, Yoshihiro Urade, Toshiji Saibara, Jude A. Oben.
  • Journal Title: Journal of Gastroenterology Volume: 49(6) Issue: 6 Pages: 1065-1173
  • DOI: 10.1007/s00535-013-0852-8
  • Peer Reviewed / Open Access
• [Presentation] 過剰発現したNF90-NF45複合体は筋サテライト細胞発生のマスター因子であるPax7の発現増加を引き起こす. (2015)
  • Author(s): 戸高寛, 樋口琢磨, 三輪武司, 森澤啓子, Lai Sylvia Chin See, 有川幹彦, 佐藤隆幸, 津田雅之, 坂本修士.
  • Organizer: Biochemistry and Molecular Biology 2015
  • Place of Presentation: 神戸ポートアイランド (神戸市)
  • Year and Date: 2015-12-01
• [Presentation] 膵臓における二本鎖RNA結合タンパク質NF90及びNF45の機能解析. (2015)
  • Author(s): 2. Lai Sylvia Chin See, 樋口琢磨, 杉山康憲, 森澤啓子, 戸高寛, 三輪武司, 津田雅之, 坂本修士.
  • Organizer: Biochemistry and Molecular Biology 2015
  • Place of Presentation: 神戸ポートアイランド (神戸市)
  • Year and Date: 2015-12-01
• [Presentation] 抗がん剤を処理したヒト白血病細胞HL-60の細胞内キナーゼの発現およびリン酸化動態の解析. (2015)
  • Author(s): 杉山康憲, 片山将一, 亀下勇, 森澤啓子, 樋口琢磨, 戸高寛, 木下英司, 木下-菊田恵美子, 小池透, 坂本修士.
  • Organizer: Biochemistry and Molecular Biology 2015
  • Place of Presentation: 神戸ポートアイランド (神戸市)
  • Year and Date: 2015-12-01
• [Presentation] 肝細胞癌における二本鎖RNA結合タンパク質によるケモカイン，CXCL5の発現制御機構. (2015)
  • Author(s): 三輪武司, 樋口琢磨, 戸高寛, 森澤啓子, Lai Sylvia Chin See, 坂本修士.
  • Organizer: Biochemistry and Molecular Biology 2015
  • Place of Presentation: 神戸ポートアイランド (神戸市)
  • Year and Date: 2015-12-01
• [Presentation] マルチPK抗体とPhos-tagを利用した細胞内リン酸化動態の新規解析法 (2015)
  • Author(s): 杉山 康憲、亀下 勇、坂本 修士
  • Organizer: 第66回日本電気泳動学会総会
  • Place of Presentation: 東京工科大学 (東京都)
  • Year and Date: 2015-09-04
  • Invited
• [Presentation] 二本鎖RNA結合タンパク質が関与する肝細胞癌の新たな遊走能制御 (2014)
  • Author(s): 樋口琢磨, 安東徳子, 三輪武司, 戸高寛, 森澤啓子, Lai Chin See Sylvia, 杉山康憲, 津田雅之, 坂本修士.
  • Organizer: 第87回 日本生化学会
  • Place of Presentation: 国立京都国際会館（京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] マルチPK抗体とPhos-tagを利用した細胞内リン酸化シグナリング解析法 (2014)
  • Author(s): 杉山康憲, 森澤啓子, 樋口琢磨, 戸高寛, 山口史佳, 安東徳子, 坂本修士.
  • Organizer: 第87回 日本生化学会
  • Place of Presentation: 国立京都国際会館（京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Identification of a motif in NF90 protein that is involved in inhibition of microRNA biogenesis. (2013)
  • Author(s): Yamaguchi F, Todaka H, Higuchi T, Morisawa K, Sugiyama Y, Sakamoto S.
  • Organizer: 第36回 日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド (神戸)
• [Presentation] NF90-NF45 complex impairs muscular maturation via blockade of miR-133 biogenesis. (2013)
  • Author(s): Todaka H, Higuchi T, Yagyu K, Yamaguchi F, Morisawa K, Fukushima A, Tsuda M, Sugiyama Y, Sakamoto S.
  • Organizer: 第36回 日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド (神戸)
• [Presentation] NF90-NF45 regulates an expression of EGFR through suppression of miR-7 biogenesis in hepatocellular carcinoma. (2013)
  • Author(s): Higuchi T, Todaka H, Morisawa K, Yamaguchi F, Ono M, Sugiyama Y, Tsuda M, Sakamoto S.
  • Organizer: 第36回 日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド (神戸)
• [Presentation] Suppression of miR-7 biogenesis by NF90-NF45 complex in Hepatocellular Carcinoma. (2013)
  • Author(s): Higuchi T, Todaka H, Yamaguchi F, Morisawa K, Ono M, Sugiyama Y, Tsuda M, Sakamoto S.
  • Organizer: 第15回RNA学会年会
  • Place of Presentation: 愛媛県県民文化会館ひめぎんホール (愛媛)
• [Presentation] Overexpression of NF90-NF45 complex causes centronuclear muscle fibers through blockade of microRNA biogenesis. (2013)
  • Author(s): Todaka H, Higuchi T, Yagyu K, Yamaguchi F, Morisawa K, Fukushima A, Tsuda M, Sugiyama Y, Sakamoto S.
  • Organizer: 第15回RNA学会年会
  • Place of Presentation: 愛媛県県民文化会館ひめぎんホール (愛媛)
• [Presentation] NF90及びNF45によるmiRNA-7生合成阻害を介した腫瘍化制御機構の解明 (2013)
  • Author(s): 樋口琢磨, 戸高寛, 池恩秀, 山口史佳, 森澤啓子, 小野正文, 杉山康憲, 津田雅之, 坂本修士, 谷口武利.
  • Organizer: 第54回日本生化学会 中国・四国支部例会
  • Place of Presentation: 徳島大学 大塚講堂 (徳島)
• [Presentation] NF90-NF45複合体によるmiRNA産生調節を介した筋成熟抑制 (2013)
  • Author(s): 戸高寛, 樋口琢磨, 矢生健一, 森澤啓子, 山口史佳, 池恩秀, 福島敦樹, 津田雅之, 杉山康憲, 坂本修士.
  • Organizer: 第54回日本生化学会 中国・四国支部例会
  • Place of Presentation: 徳島大学 大塚講堂 (徳島)