The selective collection of stress granules using nanopipette

• Project/Area Number: 15K14707
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: University of Tsukuba
• Principal Investigator: KIM Jundal 筑波大学, 生命領域学際研究センター, 助教 (90570036)
• Co-Investigator(Renkei-kenkyūsha): FUKAMIZU Akiyoshi 筑波大学, 生命領域学際研究センター, 教授 (60199172)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: ストレス顆粒 / ナノピペット

Outline of Final Research Achievements

When cells are under environmental stress, DNA and RNA can be easily damaged, which cause biological disorder. To prevent from several biological disorders, there exist a lot of self-protective systems in cells. In recent years, a stress-responsive self-protective system called stress granule (SG) has been discovered. SG is cytoplasmic aggregation that forms when translation initiation is impaired by stress, and it is considered to play a critical role in regulation of mRNA metabolism and protect RNAs from harmful conditions. However, the identity of SG is poorly understood, because the isolation of SG is extremely difficult. For further analysis, a method that can effectively isolate SG is demanded. Nanopipette is a robotic nanometer-size glass pipette. It can take samples selectively and rapidly in an individual cell, which may meet the requirements for SG isolation. In the present study, we attempted to collect SG using nanopipette, by marking SG with fluorescent protein markers.

Research Products

• [Journal Article] Angiodysplasia in embryo lacking protein arginine methyltransferase 1 (PRMT1) in vascular endothelial cells. (2017)
  • Author(s): Ishimaru, T., Ishida, J., Kim, JD., Mizukami, H., Hara, K., Hashimoto, M., Yagami, KI., Sugiyama, F., and Fukamizu, A.
  • Journal Title: Journal of Biochemistry Volume: 161 Pages: 255-258
  • DOI: 10.1093/jb/mvw095
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Novel helical assembly in arginine methyltransferase 8 (2016)
  • Author(s): Toma-Fukai S, Kim JD, Park KE, Kuwabara N, Shimizu N, Krayukhina E, Uchiyama S, Fukamizu A, Shimizu T
  • Journal Title: Journal of Molecular Biology Volume: 428 Issue: 6 Pages: 1197-1208
  • DOI: 10.1016/j.jmb.2016.02.007
  • Peer Reviewed / Open Access
• [Journal Article] PRMT8 as a phospholipase regulates Purkinje cell dendritic arborization and motor coordination. (2015)
  • Author(s): Kim, J.D., Park, K.E., Ishida, J., Kako, K., Hamada, J., Kani, S., Takeuchi, M., Namiki, K., Fukui, H., Fukuhara, S., Hibi, M,. Kobayashi, M., Kanaho, Y., Kasuya, Y., Mochizuki, N., Fukamizu, A.
  • Journal Title: Sci. Adv. Volume: 1 Issue: 11
  • DOI: 10.1126/sciadv.1500615
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] PRMT1 v2 ノックアウトマウス由来MEFを用いたスプライシングバリアントの細胞機能解析 (2016)
  • Author(s): 水上早瀬、石田純治、金 俊達、水野聖哉、杉山文博、深水昭吉
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-12-02
• [Presentation] PRMT8 as a phospholipase is required for Purkinje cell dendritic arborization and motor coordination (2016)
  • Author(s): Jun-Dal Kim、Junji Ishida、 Akiyoshi Fukamizu
  • Organizer: International meeting of the federation of Korean microbiological societies
  • Place of Presentation: KINTEX、 (Gyeonggi-do・Korea)
  • Year and Date: 2016-11-03
  • Int'l Joint Research
• [Presentation] 生体内におけるPRMT1スプライシングバリアントの機能 (2016)
  • Author(s): 水上早瀬、石田純治、金 俊達、水野聖哉、杉山文博、深水昭吉
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター（宮城県・仙台市）
  • Year and Date: 2016-09-27
• [Presentation] ナノピペットを用いた単一細胞内の下流の単離 (2016)
  • Author(s): 金 俊達、深水昭吉、ジンウイクン、トートエステル
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター（宮城県・仙台市）
  • Year and Date: 2016-09-26
• [Presentation] ホスホリパーゼ活性を持つアルギニンメチル化酵素の小脳機能に対する役割 (2016)
  • Author(s): 金 俊達、石田純治、加香孝一郎、濱田樹理、深水昭吉
  • Organizer: 第10回炎症・脂質代謝・メタボリサーチフォーラム
  • Place of Presentation: 東京大学山上会館、東京、文京区
  • Year and Date: 2016-02-13
• [Presentation] ヒスタミンのH3受容体を介した心腎病態に対する作用の検討 (2015)
  • Author(s): 野口和之、石田純治、石丸友博、川崎祥平、金 俊達、権 哲源、加香孝一郎、山縣邦弘、深水昭吉
  • Organizer: Advans 研究会2015
  • Place of Presentation: ホテルグランドパレス、東京都、千代田区
  • Year and Date: 2015-12-12
• [Presentation] 血管内皮タンパク質アルギニンメチル基転移酵素・PRMT1の機能解析 (2015)
  • Author(s): 石丸友博、石田純治、金 俊達、橋本美涼、水上早瀬、杉山文博、八神健一、深水昭吉
  • Organizer: 第19回日本心血管内分泌代謝学会学術総会
  • Place of Presentation: 神戸国際会議場、兵庫県、神戸
  • Year and Date: 2015-12-10
• [Presentation] アルギニンメチル化酵素PRMT8のホスホリパーゼ活性と生物学的意義 (2015)
  • Author(s): 金 俊達、石田純治、加香孝一郎、濱田樹理、深水昭吉
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド、兵庫県、神戸
  • Year and Date: 2015-12-02