Study on the cellular distribution and molecular function of ALS2, a product of the novel causative gene for famrlial ALS

• Project/Area Number: 14380361
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Tokai University
• Principal Investigator: HADANO Shinji Tokai University, Medical Research Institute, Associate Professor, 総合医学研究所, 助教授 (60281375)
• Co-Investigator(Kenkyū-buntansha): OSUGA Hitoshi Tokai University, Medical Research Institute, Associate Professor, 総合医学研究所, 非常勤助教授 (60203775) ; TANAKA Kazunori Tokai University, School of Medicine, Assistant Researcher, 医学部, 助手 (10338767)
• Project Period (FY): 2002 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥14,800,000 (Direct Cost: ¥14,800,000); Fiscal Year 2004: ¥4,300,000 (Direct Cost: ¥4,300,000); Fiscal Year 2003: ¥4,700,000 (Direct Cost: ¥4,700,000); Fiscal Year 2002: ¥5,800,000 (Direct Cost: ¥5,800,000)
• Keywords: amyotrophic laterel sclerosis / familial ALS / ALS2 gene / motor neuron / neurodegenerative disease / guanine nucleotide exchange factor / small GTPase / vesicle trafficking / 細胞内膜・小胞移送 / タンパク質ドメイン

Research Abstract

The aim of this study was to delineate molecular function of ALS2, a causative gene product underlying a number of recessive motor neuron diseases, thereby clarifying the molecular mechanisms for neuronal cell dysfunction and death resulted from loss of function mutations in the ALS2 gene. We have carried out the following four sets of molecular, biochemical, and cell biological studies; 1) immunocytochemical analysis of the subcellular localization of ALS2, 2) identification of the target small GTPases for ALS2, 3) identification of the ALS2 associating/binding factors, and 4) exploration of the biochemical signal pathways mediated by ALS2. We here demonstrate that ALS2 specifically binds to small GTPase Rab5 and functions as a GEF for Rab5. Ectopically expressed ALS2 localizes with Rab5 onto early endosomal compartments, and stimulates the enlargement of endosomes in the cultured neuronal and non-neuronal cells. The carboxy-terminus of ALS2 carrying the MORN/VPS9 domain mediates not only the activation of Rab5 via a guanine-nucleotide exchanging reaction, but also the endosomal localization for ALS2, thereby leading to endosome fusions by activating the Rab5-EEAI-SNARE molecular pathway. We have also shown that ALS2 forms a homophilic oligomer through its distinct C-terminal regions. This homo-oligomerization is crucial for the Rab5GEF activity and the ALS2-mediated endosome enlargement in the cells. Lastly, we have identified a novel ALS2 homologous gene, ALS2 C-terminal like (ALS2CL), which is a novel factor modulating the Rab5-mediated endosome dynamics in the cells. Taken together, a perturbation of the endosomal dynamics, which is resulted from a loss of ALS2 Rab5-GEF activity, might be a key molecular basis underlying neuronal dysfunction and degeneration in a number of motor neuron diseases caused by the ALS2 mutations.

Research Products

• [Journal Article] A Dopamine D4 receptor antagonist attenuates ischemia-induced neuronal cell damage viaupregulation of neuronal apoptosis inhibitory protein. (2005)
  • Author(s): Yoshinori Okada
  • Journal Title: J.Cereb.Blood Flow Metab. (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A Dopamine D4 receptor antagonist attenuates ischemia-induced neuronal cell damage via upregulation of neuronal apoptosis inhibitory protein. (2005)
  • Author(s): Okada, Y., Sakai, H., Kohiki, E., Suga, E., Yanagisawa, Y., Tanaka, K., Hadano, S., Osuga, H., Ikeda, J.-E.
  • Journal Title: J.Cereb.Blood Flow Metab. (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genes for amyotrophic lateral sclerosis. (2005)
  • Author(s): Hadano, S.
  • Journal Title: Yobou Igaku Jiten (Asakura Publisher) (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel nuclear shuttle proteins, HDBP1 and HDBP2, bind to neuronal cell specificcis-regulatory element in the promoter for the human Huntington's disease (2004)
  • Author(s): Kazunori Tanaka
  • Journal Title: J.Biol.Chem. 279 Pages: 7275-7286
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Homo-oligomerization of ALS2 through its unique carboxy-terminal region is essential for the ALS2-associated Rab5 guanine nucleotide exchange activity and its regulatory function on endosome trafficking. (2004)
  • Author(s): Ryota Kunita
  • Journal Title: J.Biol.Chem. 279 Pages: 38626-38635
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] ALS2CL, the novel protein highly homologous to the carboxy-terminal half of ALS2, binds to Rab5 and modulates endosome dynamics. (2004)
  • Author(s): Shinji Hadano
  • Journal Title: FEBS Lett. 575 Pages: 64-70
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Novel nuclear shuttle proteins, HDBP1 and HDBP2, bind to neuronal cell specific cis-regulatory element in the promoter for the human Huntington's disease gene. (2004)
  • Author(s): Tanaka, K., Shouguchi-Miyata, J., Miyamoto, N., Ikeda, J.-E.
  • Journal Title: J.Biol.Chem. 279 (8) Pages: 7275-7286
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] (2004) Homo-oligomerization of ALS2 through its unique carboxy-terminal region is essential for the ALS2-associated Rab5 guanine nucleotide exchange activity and its regulatory function on endosome trafficking. (2004)
  • Author(s): Kunita, R., Otomo, A., Mizumura, H., Suzuki, K., Showguchi-Miyata, J., Yanagisawa, Y., Hadano, S., Ikeda, J.-E.
  • Journal Title: J.Biol.Chem. 279 (37) Pages: 38626-38635
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] ALS2CL, the novel protein highly homologous to the carboxy-terminal half of ALS2, binds to Rab5 and modulates endosome dynamics. (2004)
  • Author(s): Hadano, S., Otomo, A., Suzuki-Utsunomiya, K., Kunita, R., Yanagisawa, Y., Showguchi-Miyata, J., Mizumura, H., Ikeda, J.-E.
  • Journal Title: FEBS Lett. 575 (1-3) Pages: 64-70
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Homo-oligomerization of ALS2 through its unique carboxy-terminal region is essential for the ALS2-associated Rab5 guanine nucleotide exchange activity and its regulatory function on endosome trafficking. (2004)
  • Author(s): Ryota Kunita
  • Journal Title: J.Biol.Chem. 279・37 Pages: 38626-38635
• [Journal Article] ALS2CL, the novel protein highly homologous to the carboxy-terminal half of ALS2, binds to Rab5 and modulates endosome dynamics. (2004)
  • Author(s): Shinji Hadano
  • Journal Title: FEBS Lett. 575・1-3 Pages: 64-70
• [Journal Article] ALS2, a novel guanine nucleotide exchange factor for the small GTPase Rab5, is implicated in endosomal dynamics. (2003)
  • Author(s): Asako Otomo
  • Journal Title: Hum.Mol.Genet. 12 Pages: 1671-1687
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Single-nucleotide polymorphisms in uncoding regions of ALS2 gene of Japanese patients with autosomal-recessive amytrophic lateral sclerosis. (2003)
  • Author(s): Isao Nagano
  • Journal Title: Neurol.Res 25 Pages: 505-509
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 新しい家族性筋萎縮性側索硬化症遺伝子:ALS2-運動ニューロン疾患における分子接点 (2003)
  • Author(s): 秦野 伸二
  • Journal Title: 医学のあゆみ 205 Pages: 119-123
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] ALS2, a novel guanine nucleotide exchange factor for the small GTPase Rab5, is implicated in endosomal dynamics. (2003)
  • Author(s): Otomo, A., Hadano, S., Okada, T., Mizumura, H., Kunita, R., Nishijima, H., Showguchi-Miyata, J., Yanagisawa, Y., Kohiki, E., Suga, E., Yasuda, M., Osuga, H., Nishimoto, T., Naurumiya, S., Ikeda, J.-E.
  • Journal Title: Hum.Mol.Genet. 12 (14) Pages: 1671-1687
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Single-nucleotide polymorphisms in uncoding regions of ALS2 gene of Japanese patients with autosomal-recessive amyotrophic lateral sclerosis. (2003)
  • Author(s): Nagano, I., Murakami, T., Shiote, M., Manabe, Y., Hadano, S., Yanagisawa, Y., Ikeda, J.-E., Abe, K.
  • Journal Title: Neurol.Res. 25 (5) Pages: 505-509
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] ALS2, the novel gene for a form of recessive amyotrophic lateral sclerosis. -implication in the pathogenesis for motor neuron diseases. (2003)
  • Author(s): Hadano, S.
  • Journal Title: Igaku no Ayumi 205 (2) Pages: 119-123
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional human NAIP promoter and transcriptional elements of the human NAIP and ΨNAIP genes. (2002)
  • Author(s): Ming Xu
  • Journal Title: Biochim.Biophys.Acta 1574 Pages: 35-50
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Unstable transmission of the RS447 human megasatellite tandem repetitive sequence that contains the USP17 gene encoding a functional deubiquitinating enzyme. (2002)
  • Author(s): Takeya Okada
  • Journal Title: Hum.Genet. 110 Pages: 302-313
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis. (2002)
  • Author(s): Roshni R.Singaraja
  • Journal Title: Hum.Mol.Genet. 11 Pages: 2815-2828
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 家族性ALSの新たな遺伝子 (2002)
  • Author(s): 秦野 伸二
  • Journal Title: BRAIN MEDICAL 14 Pages: 47-54
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] ALSの原因遺伝子 (2002)
  • Author(s): 秦野 伸二
  • Journal Title: 生化学 74 Pages: 483-489
  • NAID: 10008692567
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 筋萎縮性側索硬化症原因遺伝子の同定とゲノム研究 (2002)
  • Author(s): 秦野 伸二
  • Journal Title: ゲノム医学 2 Pages: 231-240
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Functional human NAIP promoter and transcriptional elements of the human NAIP and ΨNAIP gene (2002)
  • Author(s): Xu, M., Okada, T., Sakai, H., Miyamoto, N., Yanagisawa, Y., MacKenzie, A.E., Hadano, S., Ikeda, J.-E.
  • Journal Title: Biochim.Biophys.Acta 1574 (1) Pages: 35-50
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Unstable transmission of the RS447 human megasatellite tandem repetitive sequence that contains the USP17 gene encoding a functional deubiquitinating enzyme. (2002)
  • Author(s): Okada, T., Gondo, Y., Goto, J., Kanazawa, I., Hadano, S., Ikeda, J.-E.
  • Journal Title: Hum.Genet. 110 (4) Pages: 302-313
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis. (2002)
  • Author(s): Singaraja, R.R., Hadano, S., Metzler, M., Givan, S., Wellington, C.L, Warby, S., Yanai, A., Gutekunst, C.-A., Leavitt, B.R., Yi, H., Fichter, K., Gan, L, McCutcheon, K., Chopra, V., Michel, J., Hersch, S.M., Ikeda, J.-E., Hayden, M.R.
  • Journal Title: Hum.Mol.Genet. 11 (23) Pages: 2815-2828
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The novel gene for the familial ALS. (2002)
  • Author(s): Hadano, S.
  • Journal Title: BRAIN MEDICAL 14 (1) Pages: 47-54
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Causative genes for familial amyotrophic lateral sclerosis. (2002)
  • Author(s): Hadano, S.
  • Journal Title: Seikagaku 74 (6) Pages: 483-489
  • NAID: 10008692567
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic and genomic approaches for identification of genes underlying amyotrophic lateral sclerosis. (2002)
  • Author(s): Hadano, S.
  • Journal Title: Genome Med. 2 (3) Pages: 231-240
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The causative gene for familial ALS2. (2002)
  • Author(s): Hadano, S., Ikeda, J.-E.
  • Journal Title: Nou Kinou No Kaimei (Gaia Publisher) Pages: 299-305
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A Dopamine D4 receptor antagonist attenuates ischemia-induced neuronal cell damage via upregulation of neuronal apoptosis inhibitory protein.
  • Author(s): Yoshinori Okada
  • Journal Title: J.Cereb.Blood Flow Metab. (in press)
• [Book] 予防医学事典(松島網治, 酒井敏行, 石川昌, 稲寺秀邦編)、116.筋萎縮性側索硬化症原因遺伝子 (2005)
  • Author(s): 秦野 伸二(分担著)
  • Publisher: 朝倉書店、東京(印刷中)
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 脳機能の解明-生命科学の主潮流-(赤池紀扶, 東英穂, 阿部康二, 久保千春編)、第8章神経変性疾患の遺伝子異常と病態解明、家族性筋萎縮性側策硬化症2型(ALS2)の原因遺伝子 (2002)
  • Author(s): 秦野 伸二(分担著)
  • Publisher: ガイア出版会、福岡
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Asako Otomo: "ALS2, a novel guanine nucleotide exchange factor for the small GTPase Rab5, is implicated in endosomal dynamics"Hum.Mol.Genet.. 12・14. 1671-1687 (2003)
• [Publications] Isao Nagano: "Single-nucleotide polymorphisms in uncoding regions of ALS2 gene of Japanese patients with autosomal-recessive amyotrophic lateral sclerosis"Neurol.Res.. 25・5. 505-509 (2003)
• [Publications] Kazunori Tanaka: "Novel nuclear shuttle proteins, HDBP1 and HDBP2, bind to neuronal cell specific cis-regulatory element in the promoter for the human Huntington's disease gene"J.Biol.Chem.. (in press). (2004)
• [Publications] 秦野 伸二: "新しい家族性筋萎縮性側索硬化症遺伝子:ALS2-運動ニューロン疾患における分子接点"医学のあゆみ. 205・2. 119-123 (2003)
• [Publications] 秦野 伸二: "予防医学事典(松島綱治、酒井敏行、石川昌、稲寺秀邦編)、116.筋萎縮性側索硬化症原因遺伝子"朝倉書店、東京(印刷中). (2004)
• [Publications] Ming Xu: "Functional human NAIP promoter and transcriptional elements of the human NAIP and ψNAIP genes"Biochim. Biophys. Acta. 1574(1). 35-50 (2002)
• [Publications] Takeya Okada: "Unstable transmission of the RS447 human megasatellite tandem repetitive sequence that contains the USP17 gene encoding a functional deubiquitinating enzyme"Hum. Genet.. 110(4). 302-313 (2002)
• [Publications] Roshni R.Singaraja: "HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis"Hum. Mol. Genet.. 11(23). 2815-2828 (2002)
• [Publications] 秦野伸二: "家族性ALSの新たな遺伝子"BRAIN MEDICAL. 14(1). 47-54 (2002)
• [Publications] 秦野伸二: "ALSの原因遺伝子"生化学. 74(6). 483-489 (2002)
• [Publications] 秦野伸二: "筋萎縮性側索硬化症原因遺伝子の同定とゲノム研究"ゲノム医学. 2(3). 231-240 (2002)
• [Publications] 秦野伸二 (分担著): "脳機能の解明-生命科学の主潮流-(赤池紀扶、東英穂、阿部康二、久保千春編)、第8章神経変性疾患の遺伝子異常と病態解明、家族性筋萎縮性側索硬化症2型(ALS2)の原因遺伝子"ガイア出版会、福岡. 299-305 (2002)