植物の生存戦略における液胞機能の総合的理解

• Project/Area Number: 10219207
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Okazaki National Research Institutes
• Principal Investigator: 大隅 良典 (大隈 良典) 岡崎国立共同研究機構, 基礎生物学研究所, 教授 (30114416)
• Co-Investigator(Kenkyū-buntansha): 西村 幹夫 岡崎国立共同研究機構, 基礎生物学研究所, 教授 (80093061) ; 竹川 薫 香川大学, 農学部, 教授 (50198272) ; 安楽 泰宏 帝京科学大学, 理工学部, 教授 (20012643) ; 山本 昭平 名古屋大学, 大学院・生命農学研究科, 教授 (70210341) ; 三村 徹郎 奈良女子大学, 理学部, 教授 (20174120) ; 野田 健司 岡崎国立共同研究機構, 基礎生物学研究所, 助手 (00290908) ; 鎌田 芳彰 岡崎国立共同研究機構, 基礎生物学研究所, 助手 (20291891) ; 大隅 萬里子 帝京科学大学, 理工学部, 教授 (40168927)
• Project Period (FY): 1998 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥170,900,000 (Direct Cost: ¥170,900,000); Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2002: ¥38,300,000 (Direct Cost: ¥38,300,000); Fiscal Year 2001: ¥35,600,000 (Direct Cost: ¥35,600,000); Fiscal Year 2000: ¥31,500,000 (Direct Cost: ¥31,500,000); Fiscal Year 1999: ¥31,500,000 (Direct Cost: ¥31,500,000); Fiscal Year 1998: ¥32,500,000 (Direct Cost: ¥32,500,000)
• Keywords: オートファジー / Atg / 酵母 / ユビキチン様タンパク質 / 膜形成 / オートファゴソーム / APG / 窒素飢餓 / 蛋白質分解 / ALD6 / 液胞 / 老化 / タンパク質分解 / Apg9 / 栄養飢餓 / APG6 / VPS30 / APG14 / VPS34 / イノシトール3リン酸(Pl3P) / APG2 / 膜動態 / タンパク結合反応 / Apg / Apg8 / Apg12 / タンパク質結合反応 / 自食作用

Research Abstract

平成10年発足の特定領域研究「植物の生存戦略における液胞機能を総合的理解」(代表者、基礎生物学研究所・大隅良典)の終了を受けて、これまでの成果を広く国内外の液胞研究者と議論し、今後の領域の展望を明らかにすることを目的として本会議は開催された。国内外から予想を遙かに超える125名の参加者(外国人研究者25人を含む)を得て、非常に内容的に高い国際会議であった。ポスター発表も当初の予測を大幅に上まわりほぼ30の発表がなされ、熱心な議論が行われた。
主なる内容は、液胞形成機構とそのダイナミズム、分解コンパートメントとしての液胞機能、液胞の分化と発生、液胞膜輸送、ポンプと2次的輸送系、花弁の色素をめぐる諸問題など多岐に亘った。近年のゲノム情報の進展、細胞内タンパク質の可視化技術の進歩、遺伝生化学的な手法を背景に最先端の研究内容が紹介された。液胞膜のもつポンプ、輸送体などの解析に加えて、環境耐性、老化、プログラム細胞死、花色制御、オートファジーさらに重力屈性などの植物の持つ高次機能に液胞が深く関わっていることが認識された。研究対象もシロイヌナズナ、タバコ、アサガオ、カサノリ、酵母、麹菌など多岐に亘ったが違和感もなくその研究の本質に関する質疑が活発に行われた。これらの多様な研究が相互に関連していることの認識が広まった点が、過去に行われた液胞の国際会議と明らかに異なる特徴であった。これまで個別に研究されてきた液胞機能を総合的に捉え、様々な協力関係が生まれる契機となった点で大きな意義があったと思われる。
なお終了後、海外参加者から会議のすばらしさに関して多数の謝辞が送られてきたことも会議の成功を裏付けるものである。

Research Products

• [Publications] Mizushima, N., Kuma, A., Kobayashi, Y., Yamamoto, A., Takao, T., Natsume, T., Ohsumi, Y., Yoshimori, T.: "Mouse Apg16L, a novel WD repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate"J.Cell Sci.. 116. 1679-1688 (2003)
• [Publications] Hamasaki, M., Noda, T., Ohsumi, Y.: "The early secretory pathway contributes to autophagosome formation."Cell Struc.Func.. 28. 49-54 (2003)
• [Publications] Sugawara, K., Suzuki, N., Fujioka, Y., Mizushima, N., Ohsumi, Y., Inagaki, F.: "Crystalization and preliminary X-ray analyses of LC3-I"Acta Crys.. D59. 1464-1465 (2003)
• [Publications] Mukaiyama, H., Baba, M., Osumi, M., Aoyagi, S., Kato, N., Ohsumi, Y., Sakai, Y.: "Modification of a Ubiquitin-like protein Paz2 conducted micropexophagy through formation of a membrane structure"Mol.Biol.Cell.. (in press). (2003)
• [Publications] Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A, Rosen J, Eskelinen EL, Mizushima N, Ohsumi Y, Cattoretti G, Levine B.: "Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene"J.Clin.Invest. 112. 1809-1820 (2003)
• [Publications] Klionsky DJ, Cregg JM, Dunn WA Jr, Emr SD, Sakai Y, Sandoval IV, Sibirny A, Subramani S, Thumm M, Veenhuis M, Ohsumi Y.: "A Unified Nomenclature for Yeast Autophagy-Related Genes"Dev.Cell. 5. 539-545 (2003)
• [Publications] Mizushima, N., Yamamoto, A., Matsui, M., Yoshimori, T., Ohsumi, Y.: "In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker"Mol.Biol.Cell. (in press). (2004)
• [Publications] Ohsumi, Y.: "Cellular recycling system- molecular mechanism of autophagy"In Cell Growth, Cold Spring Harbor Press. (in press). (2003)
• [Publications] Ohsumi, Y., Mizushima, N.: "Two ubiquitin-like conjugation systems essential for autophagy"Sem.Cell Dev.Biol.. (in press). (2004)
• [Publications] 奈良, 水島, 山本, 壁谷, 大隅, 吉森: "SKD1, AAA ATPase-dependent endosomal transport is autolysosome formation"Cell Struct.Func.. 27. 29-37 (2002)
• [Publications] 野田, 鈴木, 大隅: "Autophagosome formation in yeast : De novo formation of membrane structure"Trends Cell Biol.. 12. 231-235 (2002)
• [Publications] 久万, 水島, 石原, 大隅: "Formation of the 〜350 kD Apg12-Apg5-Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast"J.Biol.Chem.. 277. 18619-18625 (2002)
• [Publications] 花岡, 野田, 白野, 加藤, 林, 柴田, 田畑, 大隅: "Leaf senescence and starvation-induced chlorosis are accelerated by the disruption of an Arabidopsis thaliana autophagy gene"Plant Physiol.. 129. 1181-1193 (2002)
• [Publications] 鈴木, 中川, 吉川, 笹川, 吉森, 大隅, 西野, 石浦, 野中: "The first molecular evidence that autophagy relates rimmed vacuole formation in chloroquin myopathy"J.Biochem.. 131. 647-651 (2002)
• [Publications] 鈴木, 鎌田, 大隅: "Studies of cargo delivery to the vacuole mediated by autophagosomes in Saccharomyces cerevisiae"Developmental Cell. 3. 815-823 (2002)
• [Publications] 水島, 吉森, 大隅: "Mouse Apg10 as an Apg12 conjugating enzyme : Analysis by the conjugation-mediated yeast two hybrid method"FEBS Lett.. 532. 450-454 (2002)
• [Publications] 水島, 久万, 小林, 山本, 高尾, 夏目, 大隅, 吉森: "Mouse Apg16L, a novel WD repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate"J.Cell Sci. (in press). (2003)
• [Publications] 水島, 吉森, 大隅: "Role of the Apg12 conjugation system in mammalian autophagy, Int."J.Biochem.Cell Biol. (in press). (2003)
• [Publications] 濱崎, 野田, 大隅: "The Early Secretory Pathway Contributes to Autophagosome Formation"Cell Struc.Func. (in press). (2003)
• [Publications] 水島, 大隅, 吉森: "Autophagosome formation in Mammalian cells"Cell Struct Funct.. 27. 421-429 (2002)
• [Publications] 大隅: "Cellular Recycling System-Molecular Mechanism of Autophagy-"Cell Growth (Cold Spring Harbour Press). (in press). (2003)
• [Publications] Kihara, A. et al: "Two distinct Vps34 PtdIns 3-kinase complexes function in autophagy and CPY sorting in Saccharomyces cerevisiae"J. Cell Biol.. 152. 519-530 (2001)
• [Publications] Kihara, A. et al: "Beclin-phsophatidylinositol 3-kinase complex functions at the trans-Golgi network"EMBO Report. 2. 330-335 (2001)
• [Publications] Mizushima, N. et al: "Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells"J. Cell Biol.. 152. 657-668 (2001)
• [Publications] Komatsu, M. et al: "The C-terminal region of an Apg7p/Cvt2p is required for homodimerization and is essential for its E1-activity and E1-E2 complex-formation"J. Biol. Chem.. 276. 9846-9854 (2001)
• [Publications] Kim, J.: "Cvt9/Gsa9 functions in sequestering selective cytosolic cargo destined for the vacuole"J. Cell. Biol.. 153. 381-396 (2001)
• [Publications] Shintani, T. et al: "Apg2p functions in autophagosome formation on the perivacuolar structure"J Biol Chem.. 276. 30452-30460 (2001)
• [Publications] Ishihara, N. et al: "Autophagosome requires specific early Sec proteins for its formation and NSF/SNARE for its fusion to the vacuole"Mol. Biol. Cell. 12. 3690-3702 (2001)
• [Publications] Suzuki, K. et al: "The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation"EMBO J.. 20. 5971-5981 (2001)
• [Publications] Ohsumi Y.: "Molecular dissection of autophagy : two ubiquitin-like systems"Nat. Rev. Mol. Cell Biol.. 2. 211-216 (2001)
• [Publications] Yoshimori,T., et al,: "SKD1, an AAA-ATPase related to the yeast Vps4p, regulates morphology and transport function of early endosomes."Mol.Biol.Cell. 11. 747-763 (2000)
• [Publications] George,M,D., et al,: "Apg5p functions in the sequestration step in the cytoplasm to vacuole targeting and macroautophagy pathways."Mol.Biol.Cell. 11. 969-982 (2000)
• [Publications] Noda,T., et al,: "Apg9/Cvt7p is an integral membrane protein required for transport vesicle formation in the Cvt and autophagy"J.Cell Biol.. 148. 465-479 (2000)
• [Publications] Furukawa,K., et al,: "A protein conjugation system in yeast with homology to biosyntheic enzyme reaction of prokaryotes."J.Biol.Chem.. 275. 7462-7465 (2000)
• [Publications] Scott,S.V., et al,: "Apg13p and Vac8p are part of a complex of phosphoproteins that are required for cytoplasm to vacuole targeting."J.Biol.Chem.. 275. 25840-25849 (2000)
• [Publications] Kamada,Y., et al,: "Tor-mediated induction of autophgay via an Apg1 protein kinase complex."J.Cell Biol.. 150. 1507-1513 (2000)
• [Publications] Kirisako,T., et al,: "Reversible modification regulates the memrane-binding state of Apg8/Aut7 essential for autophagy and the cytoplasm to vacuole targeting pathway"J.Cell Biol.. 151. 263-276 (2000)
• [Publications] Kabeya,Y., et al,: "LC3, a mammalian homologue of yeast Apg8p is processed and localized in autophagosome membranes."EMBO.J.. 19. 5720-5728 (2000)
• [Publications] Grote,E, et al,: "Geranylgeranylated SNAREs are dominant inhibitors of membrane fusion."J.Cell.Biol.. 151. 453-466 (2000)
• [Publications] Scott S.V, et al,: "Apg13p and Vac8p are part of a complex of phosphoproteins that are required for cytoplasm to vacuole targeting."J Biol Chem.. 275. 25840-25849 (2000)
• [Publications] Ichimura,Y., et al,: "Ubiquitination-like system mediates novel protein lipidation."Nature. 408. 488-492 (2000)
• [Publications] Kihara,A., et al,: "Two distinct Vps34 PtdIns 3-kinase Complexes function in autophagy and CPY sorting in Saccharomyces cerevisiae."J.Cell Biol.. 152. 519-530 (2000)
• [Publications] Kihara,A., et al.: "Beclin-Phsophatidylinositol 3-kinase complex functions at the trans-Golgi Network"EMBO Report. (in press). (2001)
• [Publications] Mizushima,N., et al.: "Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells."J.Cell Biol.. (in press). (2001)
• [Publications] Komatsu,M., et al.: "The C-terminal region of an Apg7p/Cvt2p is required for homodimerization and is essential for its E1-activity and E1-E2 complex-formation."J.Biol.Chem.. (in press). (2001)
• [Publications] Kim,J., et al.: "Cvt9/Gsa9 functions in sequestering selective cytosolic cargo destined for the vacuole.74."J.Cell.Biol.. (in press). (2001)
• [Publications] Mizushima, N. et al.: "Apg16p is required for the function of the Apg12p-Apg5p conjugate in the yeast autophagic pathway"EMBO J.. 18. 3888-3896 (1999)
• [Publications] Shintani, T. et al.: "Apg10p, a novel protein-conjugating enzyme essential for autophagy in yeast"EMBO J.. 18. 5234-5241 (1999)
• [Publications] Kirisako, T. et al.: "Formation process of autophagosome is traced with Apg8/Aut7p in yeast"J. Cell Biol.. 147. 435-446 (1999)
• [Publications] George, M, D. et al.: "Apg5p functions in the sequestration step in the cytoplasm to vacuole targeting and macroautophagy pathways"Mol. Biol. Cell. (in press).
• [Publications] Noda, T. et al.: "Apg9/Cvt7p is an integral membrane protein required for transport vesicle formation in the Cvt and autophagy pathways"J. Cell Biol.. (in press).
• [Publications] Furukawa, K. et al.: "A protein conjugation system in yeast with homology to biosyntheic enzyme reavion of prokarvotes"J. Biol. Chem.. (in press).
• [Publications] Noda,T.et al,: "Tor,a phosphatidylinositol kinase homologue,controls autophagy in yeast." J.Biol.Chem.273. 3963-3966 (1998)
• [Publications] Kametaka,S.et al,: "Apg14p and Apg6/Vps30p form a protein complex essential for autophagy in the yeast, Saccharomyces cerevisiae." J.Biol.Chem.273. 22284-22291 (1998)
• [Publications] Mizushima,N.et al,: "A novel protein conjugation system essential for autophagy." Nature. 395. 395-398 (1998)
• [Publications] Mizushima,N.et al,: "A New Protein Conjugation System in Human.The counterpart of the yeast Apg12p conjugation sytem essential for autophagy." J.Biol.Chem.273. 33889-33892 (1998)
• [Publications] Klionsky,D.J.et al,: "Vacuolar import of proteins and organelles from the cytoplasm" Ann.Rev.Cell Devlop.Biol.12(in press). (1999)