Analysis of neuronal cell death by ALS-associated misfolded protein aggregation with modulation of cellular RNA homeostasis

• Project/Area Number: 23770215
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: Hokkaido University
• Principal Investigator: KITAMURA Akira 北海道大学, 先端生命科学研究科(研究院), 助教 (10580152)
• Project Period (FY): 2011 – 2014
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: ALS / タンパク質凝集体 / 神経変性疾患 / 細胞内封入体 / RNA / TDP43 / 蛍光相関分光法 / RNA結合タンパク質 / 神経細胞死 / 蛍光分光イメージング / タンパク質品質管理

Outline of Final Research Achievements

TAR-RNA/DNA binding protein 43 kDa is a disease gene product associated with amyotrophic lateral sclerosis (ALS), a motor neuron disease. In this research, fluorescence correlation spectroscopy (FCS) measurement in cell lysate revealed that a 25 kDa carboxyl-terminal fragment of TDP43 (TDP25) formed aggregation after dissociation of RNA. Next, TDP43 was sequestrated into cytoplasmic inclusion bodies of TDP25 in living cells. Neuronal cells expressing TDP25 showed higher cell death efficiency than that of TDP43. These results suggest that TDP25 may be an aggregation-causative and toxic seed, and RNA may play an important role for inhibition of aggregation-formation of TDP25.

Research Products

• [Journal Article] Moyamoya disease-associated protein mysterin/RNF213 is a novel AAA+ AT Pase, which dynamically changes its oligomeric state (2014)
  • Author(s): Daisuke Morito, Kouki Nishikawa, Jun Houseki, Akira Kitamura, Yuri Kotani, Kazumi Kiso, Masataka Kinjo, Yoshimori Fujiyoshi & Kazuhiro Nagata
  • Journal Title: SCIENTIFIC REPORTS Volume: Vol. 4 Issue: 1 Pages: 4442-4442
  • DOI: 10.1038/srep04442
  • Peer Reviewed / Open Access
• [Journal Article] Dysregulation of the Proteasome Enhances the Toxicity of ALS-linked Mutant SOD1 (2014)
  • Author(s): Kitamura, A., Inada, N., Kubota, H., Matsumoto, G., Kinjo, M., Morimoto, R.I., Nagata, K
  • Journal Title: Genes to Cells Volume: 3 Issue: 3 Pages: 209-224
  • DOI: 10.1111/gtc.12125
  • Peer Reviewed / Open Access
• [Journal Article] Prefoldin prevents aggregation of alpha-synuclein. (2014)
  • Author(s): Takano M., Tashiro E., Kitamura A., Maita H., Iguchi-Ariga M.M.S., Kinjo M., Ariga H.
  • Journal Title: Brain Research Volume: 1542 Pages: 186-194
  • DOI: 10.1016/j.brainres.2013.10.034
  • NAID: 120005397867
  • Peer Reviewed
• [Journal Article] The interaction of Hsp104 with yeast prion Sup35 as analyzed by fluorescence cross-correlation spectroscopy.oligomeric state. (2013)
  • Author(s): Ohta S., Kawai-Noma S., Kitamura A., Pack C.G., Kinjo M., Taguchi H.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 442 Issue: 1-2 Pages: 28-32
  • DOI: 10.1016/j.bbrc.2013.10.147
  • Peer Reviewed
• [Journal Article] Prefoldin protects neuronal cells from polyglutamine toxicity by preventing aggregation formation. (2013)
  • Author(s): Tashiro E, Zako T, Muto H, Itoo Y, Sorgjerd K, Terada N, Abe A, Miyazawa M, Kitamura A, Kitaura H, Kubota H, Maeda M, Momoi T, Iguchi-Ariga SM, Kinjo M, Ariga H.
  • Journal Title: Journal of Biological Chemistry Volume: 288 Issue: 27 Pages: 19958-19972
  • DOI: 10.1074/jbc.m113.477984
  • NAID: 120005323603
  • Peer Reviewed
• [Journal Article] Nonmuscle myosin II folds into a 10S form via two portions of tail for dynamic subcellular loacalization. (2013)
  • Author(s): Kiboku T.
  • Journal Title: Genes to Cells. Volume: 18(2) Issue: 2 Pages: 90-109
  • DOI: 10.1111/gtc.12021
  • Peer Reviewed
• [Journal Article] Direct association of unfolded proteins with mammalian ER stress sensor, IRE1 beta. (2012)
  • Author(s): Oikawa D.
  • Journal Title: PLoS One. Volume: 7(12) Issue: 12 Pages: e51290-e51290
  • DOI: 10.1371/journal.pone.0051290
  • Peer Reviewed
• [Journal Article] Atg9 vesicles are an important membrane source during early steps of autophagosome formation. (2012)
  • Author(s): Yamamoto H.
  • Journal Title: J. Cell Biol. Volume: 198(2) Issue: 2 Pages: 219-233
  • DOI: 10.1083/jcb.201202061
  • Peer Reviewed
• [Journal Article] Analyzing the aggregation of polyglutamine-expansion proteins and its modulation by molecular chaperones. (2011)
  • Author(s): Kubota H, Kitamura A, Nagata K.
  • Journal Title: Methods Volume: 53 Pages: 267-274
  • Peer Reviewed
• [Presentation] Characterization of cytoplasmic and nuclear inclusion of ALS-causative TDP43 protein (2011)
  • Author(s): Akira Kitamura, Yusaku Nakayama, Masataka Kinjo
  • Organizer: 第63回 日本細胞生物学会大会
  • Place of Presentation: 北海道大学（札幌市）
• [Presentation] Aggregation and disaggregation of aggregate-prone proteins: Which is toxic? (2011)
  • Author(s): Akira Kitamura
  • Organizer: 奈良先端大シンポジウム(招待講演)
  • Place of Presentation: 奈良先端大学院大学（奈良県）
• [Presentation] RNA inhibits the toxicity of carboxyl-terminal fragments of ALS-causative TDP43 protein (2011)
  • Author(s): Akira Kitamura, Yusaku Nakayama, Masataka Kinjo
  • Organizer: Cold Spring Harbor Asia, "Protein Homeostasis in Health & Disease"
  • Place of Presentation: Cold Spring Harbor Asia （中国）
• [Presentation] Characterization of cytoplasmic and nuclear inclusion of ALS-causative TDP43 protein (2011)
  • Author(s): Akira Kitamura, Yusaku Nakayama, Masataka Kinjo
  • Organizer: 特定領域「タンパク質の社会」若手シンポジウム
  • Place of Presentation: アクティプラザ琵琶（滋賀県）
• [Presentation] プロテアソーム活性阻害からの回復過程における 細胞毒性発現機構の解明
  • Author(s): 北村朗，永田和宏，金城政孝
  • Organizer: 2013年度若手イメージング研究会
  • Place of Presentation: 東京理科大学野田キャンパス（千葉県）
• [Presentation] プロテアソーム活性阻害からの回復過程における 細胞毒性発現機構の解明
  • Author(s): 北村朗，永田和宏，金城政孝
  • 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（神戸市）
• [Presentation] Increased toxicity and aggregate formation of carboxyl-terminal fragments of ALS-linked TDP43 protein in cytoplasm.
  • Author(s): Kitamura A., Nakayama Y. Shibasaki A., & Kinjo M.
  • Organizer: FASEB summer research conference
  • Place of Presentation: Vermont Academy (USA)
• [Presentation] ALS関連TDP43タンパク質のカルボキシル末端断片の細胞質における凝集体形成と毒性はRNAによって防御されている
  • Author(s): 北村朗，柴崎愛，中山祐作，金城政孝
  • Organizer: 第35回 日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場（福岡県）