1996 年度実績報告書

植物における空胞系(Vacuolar Syotem)の動態の分子機構

研究課題

研究課題/領域番号	08304044
応募区分	総合
研究機関	岡崎国立共同研究機構
研究代表者	大隅良典岡崎国立共同研究機構, 基礎生物学研究所, 教授 (30114416)
研究分担者	三村徹郎一橋大学, 商学部, 助教授 (20174120) 前島正義名古屋大学, 農学部, 助教授 (80181577) 野口哲子奈良女子大学, 理学部, 教授 (00135823) 中村研三名古屋大学, 農学部, 教授 (80164292) 中野明彦東京大学, 大学院・理学研究科・生物科学, 助教授 (90142140)
キーワード	植物細胞 / 酵母 / 空胞系 / 小胞体 / ゴルジ体 / 液胞 / タンパク質のソーティング
研究概要	今年度から2年間行われる「植物細胞における空胞系の膜動態の分子機構」はこの分野の研究者を糾合して、研究に関する相互理解と共同研究を通じて研究の一層の発展を意図するものである。本年度は11月に班会議を行い、研究発表を通じて活発な交流をすることができ有意義なものであった。班員相互の共同研究も既に複数開始している。今年度得られた班員の研究成果は以下に要約される。野口は植物細胞で初めて小胞体、ゴルジ体間の逆行輸送を存在を証明し、この過程への微小管の関与を示した。中村は植物液胞へのタンパク質の輸送機構を解析し、移行シグナルの性質、輸送過程のオルガネラに存在するV-ATPaseの関与を初めて示すことに成功した。前島は、水チャネル、H^+-PPase,Ca^<2+>/H^+対向輸送体の3つの液胞膜輸送体の分子構造と制御機構を解析した。三村は細胞質、液胞内のリン酸濃度を測定し、細胞膜リン酸輸送活性の調節について新しい知見を得た。西村は高等植物に存在する2種の液胞が相互に変換すること、それぞれの機能発現に特異的プロセシング酵素が重要な役割を担うことを示した。森山はホヤの血中に巨大かつ超低pHの液胞を見いだし、その性質を解析した。森安は、タバコ培養細胞を用いて、蔗糖飢餓によって誘導される蛋白分解の機序を解析し、植物細胞の自食作用に関する新しい知見を得た。竹川は分裂酵母の液胞タンパクの輸送シグナル、糖鎖付加に関与する酵素の変異体の取得、ナイスタチンを用いた液胞形態異常株を取得し、それらの解析を進めた。和田は、酵母をモデル系として液胞構築に必須な遺伝子の機能を明らかにした。大隅は自食作用に必須の遺伝子群の構造と機能の解析を進めた。中野は小胞体-ゴリジ体間の逆輸送系の分子機構に関わる分子を明らかにした。

研究成果
(36件)

すべてその他

すべて文献書誌 (36件)

[文献書誌] Scott,S.V.et al,: "Cytoplasm-to vacuole targeting and autophagy employ the same machinery to deliver proteins to the yeast vacuole" Proc.Natl.Acad.Sci.USA. 98. 12304-12308 (1996)
[文献書誌] Kametaka,S.et al,: "Structural and function analyses of APG5,a gene involved in autophagy in yeast." Gene. 178. 139-143 (1996)
[文献書誌] Nakamura,N et al,: "Acidification of vacuoles is required for autophagic degradation in the yeast,Saccharomyces cerevisiase." J.Biochem.193 (in press). (1997)
[文献書誌] Funakoshi,T.et al,: "Analyses of APG13 gene involved in the autophagy in yeast,Saccharomyces cerevisiae." Gene. 179 (in press). (1997)
[文献書誌] Tabuchi,M.et al,: "The Schizosaccharomyces pombe gms1+gene encodes an UDP galactose transporter homologue required for protein galactosylation" Biochem.Biophys.Res.Commun.(in press) (1997)
[文献書誌] Sato,M.et al,: "Endoplasmic reticulum localization of Sec12p is achieved by two mechanisms." J.Cell Biol.134. 279-293 (1996)
[文献書誌] Yamanushi,T. et al,: "Characterization of yeast sarl temperature-sensitive mutants, which are defective in protein trasport from the endoplasmic reticulum" J.Biochem.120. 452-458 (1996)
[文献書誌] Nakano,A. et al,: "Identification and characterization of extragenic suppressors of the yeast sec12 ts mutation." J.Biochem.120. 642-646 (1996)
[文献書誌] Ueda,T.et al,: "An Arabidopsis gene isolated by a novel method for detecting genetic interaction in yeast encodes the GDP dissociation inhibitor of Ara4 GTPase." Plant Cell. 8. 2079-2091 (1996)
[文献書誌] Yoshioka,Y.et al,: "Transient gene expression in plant cells mediated by Agrobacterium tumefaciens : application for the analysis of vinulence loci." Plant Cell Physiol.37. 782-789 (1996)
[文献書誌] Matsuoka,K.et al,: "A vacuolar-type H+-ATPase in non-vacuolar organelle is required for the sorting of soluble vacuolar protein precursors in tobacco cells." Plant Cell. (in press). (1997)
[文献書誌] Sato,Y et al,: "A tobacco cDNA encoding a homologue of N-ethylmaleimide sensitive fusion protein (NSF) from tobacco." Plant Physiol.(in press). (1997)
[文献書誌] Koide,Y.et al,: "The N-terminal propeptide of the precursor to sporamin acts as a vacuolar targeting signal even at the C-terminus of the mature part in tobacco cells." Plant Physiol.(in press).
[文献書誌] Hatano,K.et al,: "A rapid increase in the level of binding protein (BiP) is accompanied by synthesis and degradation of storage proteins in pumpkin cotyledons." Plant Cell Physiol.38 (in press). (1997)
[文献書誌] Inoue,K.et al,: "Expression of pumpkin seed TIP (tonoplast intrinsic protein) in yeast cells." Plant Cell Physiol.38 (in press). (1997)
[文献書誌] Hiraiwa,N.et al,: "An aspartic proteinase is involved in the maturation of storage proteins in concert with the vacuolar processin enzyme" Eur.J.Biochem.(in press). (1997)
[文献書誌] Hiraiwa,N.et al,: "Expression and activation of the vacuolar processing enzyme in Saccharomyces cerevisiae." Plant J.(in press). (1997)
[文献書誌] Ehara,M.et al,: "Uptake of neutral red by the vacuoles of a green alga Micrasteias pinnatifida." Plant Cell Physiol.37. 734-741 (1996)
[文献書誌] Ueda,K.et al,: "The number and the distribution pattern of Golgi bodies in cells of Micrasterias examined by fluorescence microscopy and by electron mictoscopy." Phycological Research. (in press). (1997)
[文献書誌] Robinson,D.G.et al,: "Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons." Planta.198. 95-103 (1996)
[文献書誌] Matsumoto,H.et al,: "Response of the plant root to aluminium stress : Analysis of the inhibition of the root elongation and changes in membrane function." J.Plant Res.109. 99-105 (1996)
[文献書誌] Maeshima,M.et al,: "Proton pumps of vacuolar membrane in growing plant cell" J.Plant Res.109. 119-125 (1996)
[文献書誌] Higuchi,T.et al,: "Complete nucleotide sequence of a cDNA for an intrinsic protein in vacuolar membranes from radish (Raphanus sativus) roots (PGR96-045)." Plant Physiol.111. 947 (1996)
[文献書誌] Mimura,T.et al,: "Studies on distribution,re-translocation and homeostasis of inorganic phosphate in barley leaves." Plant Cell & Environment. 193. 113-120 (1996)
[文献書誌] Beilby M.J.et al,: "Perfusion of charophyte cells : A critical analysis of the method." J.Experimental Botany. (in press). (1997)
[文献書誌] Mimura,T.et al,: "Efficient callus initiation from leaf of mangrove plant Bruguiera sexangula in amino acid medium : Effect of NaCl on callus initiation." Journal of Plant Research. (in press). (1997)
[文献書誌] Mimura,T.et al,: "NaCl-dependent growth,ion content and regeneration of calluses initiated from the mangrove plant,Bruguiera sexangula." Journal of Plant Research. (in press). (1997)
[文献書誌] Moriyasu,Y.et al,: "Autophagy in tobacco suspension-cultured cells in response to sucrose starvarion." Plant Physiology. 111. 1233-1241 (1996)
[文献書誌] Hayashi,M.et al,: "Rapid increase in number of vanadocytes in the vanadium rich ascidian,Ascidia sydneiensis samea upon treatment of live animals with ammonium chloride." J.Exp.Zool.27. 1-7 (1996)
[文献書誌] Moriyama,Y.et al,: "Vesicular monoamine transporter in microvesicles from bovine posterior pituitaries is immunologically similar o but distinct from the ohromaffin granule counterpart in its sensitivities to 1-methyl-4-phenylpyridinium and histamine" Biochem.Biophys.Res.Commun.221. 790-794 (1996)
[文献書誌] Yamada,H.et al,: "The L-type Ca2+ channel is involved in microvesicle-mediated glutamate exocytosis from pinealycytes." J.Pineal Research. 21. 165-174 (1996)
[文献書誌] Nakamura,I.et al,: "Failure of bone resorption in osteocleroic (oc/oc) mice i possibly due to the dissociation of vacuolar proton ATPase and cytoskeleton in osteoclasts." FFBS Letter. 401. 207-212 (1997)
[文献書誌] Wada,Y.et al,: "Mutational analysis of Vam4/Ypt7p function in the vacuolar biogenesis and morphogenesis in the yeast,Saccharomyces cerevisiae." Protoplasma. 191. 126-135 (1996)
[文献書誌] Shirahama,K.et al,: "Vacuolar function in the phosphate homeostasis of the yeast Saccharomyces cerevisiae." Plant Cell Physiol.37. 1090-1093 (1996)
[文献書誌] Sato,M.et al,: "Universal template plasmid for introduction of the triple HA-epitope sequence into the cloned genes." BioTechniquies. (in press). (1997)
[文献書誌] Nakamura,N.et al,: "Vam2/Vps41p and Vam6/Vps39p are components of a protein complex on the vacuolar membrane and involved in the vacuolar assembly in the yeast Saccharomyces cerevisiae" J.Biol.Chem.(in press). (1997)

1996 年度 実績報告書

植物における空胞系(Vacuolar Syotem)の動態の分子機構

研究代表者

大隅 良典 岡崎国立共同研究機構, 基礎生物学研究所, 教授 (30114416)

研究成果

[文献書誌] Scott,S.V.et al,: "Cytoplasm-to vacuole targeting and autophagy employ the same machinery to deliver proteins to the yeast vacuole" Proc.Natl.Acad.Sci.USA. 98. 12304-12308 (1996)

[文献書誌] Kametaka,S.et al,: "Structural and function analyses of APG5,a gene involved in autophagy in yeast." Gene. 178. 139-143 (1996)

[文献書誌] Nakamura,N et al,: "Acidification of vacuoles is required for autophagic degradation in the yeast,Saccharomyces cerevisiase." J.Biochem.193 (in press). (1997)

[文献書誌] Funakoshi,T.et al,: "Analyses of APG13 gene involved in the autophagy in yeast,Saccharomyces cerevisiae." Gene. 179 (in press). (1997)

[文献書誌] Tabuchi,M.et al,: "The Schizosaccharomyces pombe gms1+gene encodes an UDP galactose transporter homologue required for protein galactosylation" Biochem.Biophys.Res.Commun.(in press) (1997)

[文献書誌] Sato,M.et al,: "Endoplasmic reticulum localization of Sec12p is achieved by two mechanisms." J.Cell Biol.134. 279-293 (1996)

[文献書誌] Yamanushi,T. et al,: "Characterization of yeast sarl temperature-sensitive mutants, which are defective in protein trasport from the endoplasmic reticulum" J.Biochem.120. 452-458 (1996)

[文献書誌] Nakano,A. et al,: "Identification and characterization of extragenic suppressors of the yeast sec12 ts mutation." J.Biochem.120. 642-646 (1996)

[文献書誌] Ueda,T.et al,: "An Arabidopsis gene isolated by a novel method for detecting genetic interaction in yeast encodes the GDP dissociation inhibitor of Ara4 GTPase." Plant Cell. 8. 2079-2091 (1996)

[文献書誌] Yoshioka,Y.et al,: "Transient gene expression in plant cells mediated by Agrobacterium tumefaciens : application for the analysis of vinulence loci." Plant Cell Physiol.37. 782-789 (1996)

[文献書誌] Matsuoka,K.et al,: "A vacuolar-type H+-ATPase in non-vacuolar organelle is required for the sorting of soluble vacuolar protein precursors in tobacco cells." Plant Cell. (in press). (1997)

[文献書誌] Sato,Y et al,: "A tobacco cDNA encoding a homologue of N-ethylmaleimide sensitive fusion protein (NSF) from tobacco." Plant Physiol.(in press). (1997)

[文献書誌] Koide,Y.et al,: "The N-terminal propeptide of the precursor to sporamin acts as a vacuolar targeting signal even at the C-terminus of the mature part in tobacco cells." Plant Physiol.(in press).

[文献書誌] Hatano,K.et al,: "A rapid increase in the level of binding protein (BiP) is accompanied by synthesis and degradation of storage proteins in pumpkin cotyledons." Plant Cell Physiol.38 (in press). (1997)

[文献書誌] Inoue,K.et al,: "Expression of pumpkin seed TIP (tonoplast intrinsic protein) in yeast cells." Plant Cell Physiol.38 (in press). (1997)

[文献書誌] Hiraiwa,N.et al,: "An aspartic proteinase is involved in the maturation of storage proteins in concert with the vacuolar processin enzyme" Eur.J.Biochem.(in press). (1997)

[文献書誌] Hiraiwa,N.et al,: "Expression and activation of the vacuolar processing enzyme in Saccharomyces cerevisiae." Plant J.(in press). (1997)

[文献書誌] Ehara,M.et al,: "Uptake of neutral red by the vacuoles of a green alga Micrasteias pinnatifida." Plant Cell Physiol.37. 734-741 (1996)

[文献書誌] Ueda,K.et al,: "The number and the distribution pattern of Golgi bodies in cells of Micrasterias examined by fluorescence microscopy and by electron mictoscopy." Phycological Research. (in press). (1997)

[文献書誌] Robinson,D.G.et al,: "Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons." Planta.198. 95-103 (1996)

[文献書誌] Matsumoto,H.et al,: "Response of the plant root to aluminium stress : Analysis of the inhibition of the root elongation and changes in membrane function." J.Plant Res.109. 99-105 (1996)

[文献書誌] Maeshima,M.et al,: "Proton pumps of vacuolar membrane in growing plant cell" J.Plant Res.109. 119-125 (1996)

[文献書誌] Higuchi,T.et al,: "Complete nucleotide sequence of a cDNA for an intrinsic protein in vacuolar membranes from radish (Raphanus sativus) roots (PGR96-045)." Plant Physiol.111. 947 (1996)

[文献書誌] Mimura,T.et al,: "Studies on distribution,re-translocation and homeostasis of inorganic phosphate in barley leaves." Plant Cell & Environment. 193. 113-120 (1996)

[文献書誌] Beilby M.J.et al,: "Perfusion of charophyte cells : A critical analysis of the method." J.Experimental Botany. (in press). (1997)

[文献書誌] Mimura,T.et al,: "Efficient callus initiation from leaf of mangrove plant Bruguiera sexangula in amino acid medium : Effect of NaCl on callus initiation." Journal of Plant Research. (in press). (1997)

[文献書誌] Mimura,T.et al,: "NaCl-dependent growth,ion content and regeneration of calluses initiated from the mangrove plant,Bruguiera sexangula." Journal of Plant Research. (in press). (1997)

[文献書誌] Moriyasu,Y.et al,: "Autophagy in tobacco suspension-cultured cells in response to sucrose starvarion." Plant Physiology. 111. 1233-1241 (1996)

[文献書誌] Hayashi,M.et al,: "Rapid increase in number of vanadocytes in the vanadium rich ascidian,Ascidia sydneiensis samea upon treatment of live animals with ammonium chloride." J.Exp.Zool.27. 1-7 (1996)

[文献書誌] Yamada,H.et al,: "The L-type Ca2+ channel is involved in microvesicle-mediated glutamate exocytosis from pinealycytes." J.Pineal Research. 21. 165-174 (1996)

[文献書誌] Nakamura,I.et al,: "Failure of bone resorption in osteocleroic (oc/oc) mice i possibly due to the dissociation of vacuolar proton ATPase and cytoskeleton in osteoclasts." FFBS Letter. 401. 207-212 (1997)

[文献書誌] Wada,Y.et al,: "Mutational analysis of Vam4/Ypt7p function in the vacuolar biogenesis and morphogenesis in the yeast,Saccharomyces cerevisiae." Protoplasma. 191. 126-135 (1996)

[文献書誌] Shirahama,K.et al,: "Vacuolar function in the phosphate homeostasis of the yeast Saccharomyces cerevisiae." Plant Cell Physiol.37. 1090-1093 (1996)

[文献書誌] Sato,M.et al,: "Universal template plasmid for introduction of the triple HA-epitope sequence into the cloned genes." BioTechniquies. (in press). (1997)

[文献書誌] Nakamura,N.et al,: "Vam2/Vps41p and Vam6/Vps39p are components of a protein complex on the vacuolar membrane and involved in the vacuolar assembly in the yeast Saccharomyces cerevisiae" J.Biol.Chem.(in press). (1997)

1996 年度実績報告書

大隅良典岡崎国立共同研究機構, 基礎生物学研究所, 教授 (30114416)