2000 Fiscal Year Annual Research Report

細胞内情報伝達機構網の研究

Research Project

Project/Area Number	09NP0601
Research Institution	Kobe University
Principal Investigator	西塚泰美神戸大学, 名誉教授 (10025546)
Co-Investigator(Kenkyū-buntansha)	米澤一仁神戸大学, バイオシグナル研究センター, 教授 (70283900) 斎藤尚亮神戸大学, バイオシグナル研究センター, 教授 (60178499) 吉川潮神戸大学, バイオシグナル研究センター, 教授 (40150354) 小野功貴神戸大学, バイオシグナル研究センター, 教授 (10243297) 深見泰夫神戸大学, 遺伝子実験施設, 助教授 (00156746)
Keywords	プロテインキナーゼC / 脂質メッセンジャー / ターゲッティング / PKN / CG-NAP / アミノ酸バランス / mTOR / チロシンキナーゼ
Research Abstract	平成12年度に以下の成果をあげた。 1 プロテインキナーゼC(PKC)と情報クロストーク機構について (1)PKCδ分子種のチロシンリン酸化の誘導におけるチロシンキナーゼの働き、(2)質量分析法によるPKCδ分子種のチロシンリン酸化部位の決定、(3)PKCδ分子種のチロシンリン酸化を介する活性制御、(4)PKCδ分子種を介するアポトーシス誘導とPKBとの拮抗作用、について明らかにした。 2 脂質メッセンジャーとPKCのターゲッティング機構について (1)phospholipase A2によって産生された脂肪酸やリゾリン脂質のPKCターゲッティングへの関与、(2)基質蛋白質のリン酸化におけるPKCターゲッティングの必要性、(3)脂質メッセンジャー(アラキドン酸、セラミドなど)によるPKCターゲッティングと活性化機構、(4)ジアシルグリセロールキナーゼ(DGK)のターゲッティングによるPKCシグナリングの制御機構、を明らかにした。 3 PKC関連リン酸化酵素と細胞機能の調節について (1)PKNと相互作用する蛋白質として、CG-NAP(centrosome and Golgi localized PKN-associated protein)を同定した。また、PKNが神経特異的転写因子NDRF/NeuroD2、癌遺伝子産物E6蛋白質に結合し、機能を制御することを見出した。(2)新規のPKNファミリーであるPKNβを同定した。(3)細胞周期制御におけるPKNの機能解析を行った。 4 病態におけるPKCシグナリングの解析について、 (1)アミノ酸バランスによるp70S6キナーゼの活性制御機構、(2)アミノ酸-mTORシグナル系とPKCの関連、を明らかにした。 5 ターゲッティングと細胞周期の調節 (1)A431細胞におけるShcおよびc-Src活性化とトランスロケーション、(2)受精におけるSrc型チロシンキナーゼの役割、を明らかにした。

Research Products

(38 results)

All Other

All Publications (38 results)

[Publications] Konishi,H.: "Tyrosine phosphorylation sites of protein kinase C δ in the H_2O_2-treated cells and the regulation of its catalytic activity in vitro."Proc.Natl.Acad.Sci.USA. (印刷中).
[Publications] Yamamoto,T.: "H_<>O_<>-induced tyrosine phosphorylation of protein kinase C δ by a mechanism independent of inhibition of protein-tyrosine phosphatase in CHO and COS-7 cells."Biochem.Biophys.Res.Commun.. 273・3. 960-966 (2000)
[Publications] Okuno,S.: "Studies on the phosphorylation of protein kinase B by Ca^<2+>/calmodulin-dependent protein kinases."J.Biochem.. 127・6. 965-970 (2000)
[Publications] Kitaura,J.: "Akt-dependent cytokine production in mast cells."J.Exp.Med.. 192・5. 729-740 (2000)
[Publications] Oka,M.: "Regulation of melanogenesis through phosphatidylinositol 3-kinase-Akt pathway in human G361 melanoma cells."J.Invest.Dermatol.. 115・4. 699-703 (2000)
[Publications] Oyasu,M: "Immunocytochemical localization of a neuron-specific thrombospondin-1-like protein, NELL2 : Light and electron microscopic studies in the rat brain."Mol.Brain Res.. 76. 151-160 (2000)
[Publications] Sumioka,K.: "Induction of 55 kDa PKN cleavage product by ischemia/reperfusion model in the rat retina."Invest.Opthal.Vis.Sci.. 41・1. 29-35 (2000)
[Publications] Sakai,N.: "Involvement of the actin cytoskeleton in the regulation of serotonin transporter (SET) activity : possible mechanism underlying SET regulation by protein kinase C."Neurochem.Int.. 36. 567-579 (2000)
[Publications] Shirai,Y.: "Phospholipase A_2 and its products are involved in the purinergic receptor-mediated translocation of protein kinase C in CHO-K1 cells."J.Cell Sci.. 113. 1335-1343 (2000)
[Publications] Akiyama,H.: "Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain."Brain Res.. 858. 172-176 (2000)
[Publications] Tanii,H.: "Involvement of GABA neurons in allylnitrile-induced dyskinesia."Brain Res.. 887. 454-459 (2000)
[Publications] Ueyama,T.: "cDNA cloning of an alternative splicing variant of protein kinase Cδ(PKCδIII), a new truncated form of PKCδ, in rats."Biochem.Biophys.Res.Commun.. 269・2. 557-563 (2000)
[Publications] Shirai,Y.: "Subtype-specific translocation of diacylglycerol kinase α and γ and its correlation with protein kinase C."J.Biol.Chem.. 275・32. 24760-24766 (2000)
[Publications] Nakagawa,N.: "ENH, containing PDZ and LIM domain, heart/skeletal muscle-specific protein, associates with cytoskeletal protein through its PDZ domain."Biochem.Biophys.Res.Commun.. 272. 505-512 (2000)
[Publications] Shibaguchi,H.: "Role of synaptophysin in exocytic release of dopamine from Xenopus oocytes injected with rat brain mRNA."Cellular and Molecular Neurobiology. 20. 401-408 (2000)
[Publications] Ohmori,S.: "Importance of PKC targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS)."J.Biol.Chem.. 275・34. 26449-26457 (2000)
[Publications] Larsen,E.C.: "Differential requirement for classic and novel PKC isoforms in respiratory burst and phagocytosis in RAW 264.7 cells."J.Immunol.. 165. 2809-2817 (2000)
[Publications] Yokogawa,T.: "Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface : use of immunostaining technique with monoclonal antibodies specific for PI 3,4-P_2."FEBS Lett.. 473. 222-226 (2000)
[Publications] Kondoh,T.: "In vivo gene transfer into periventricular region by electroporation."Neurologia medico-chirurgica.. 40. 618-623 (2000)
[Publications] Kajimoto,T.: "Subtype-specific translocation of the d subtype of protein kinase C and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells."Mol.Cell.Biol.. 21. 1769-1783 (2001)
[Publications] Nakai,M.: "Amyloid β protein activates PKC-δ and induces translocation of myristoylated alanine-rich C kinase substrate (MARCKS) in microglia."Neurochem.Int.. (印刷中).
[Publications] Mishima,K.: "Molecular mechanism for α2-adrenoceptor-mediated regulation of synoviocytes proliferation."Jpn.J.Pharmacol.,. (印刷中).
[Publications] Yonezawa,K: "Nutritional regulation of gene and protein expression."Curr.Opin.Clinic.Nutri.Metab.Care. 3. 253-254 (2000)
[Publications] Takahashi,T.: "Carboxyl-terminal region conserved among phosphoinositide-kinase-related kinases is indispensable for mTOR function in vivo and in vitro."Genes to Cells. 5. 765-775 (2000)
[Publications] Sato K.-I.: "Fertilization signalling and protein-tyrosine kinases."Comp.Biochem.Physiol.B.. 126. 129-148 (2000)
[Publications] Sato K.-I.: "Regulation of Xenopus oocytes, eggs, and embryos by protein-tyrosine phosphorylation."Curr.Top.Biochem.Res.. 2. 91-102 (2000)
[Publications] Sato K.-I.: "Tyrosine kinase-dependent activation of phospholipase Cγis required for calcium transient in Xenopus egg fertilization."Dev.Biol.. 224. 453-469 (2000)
[Publications] Sato K.-I.: "Adaptor protein Shc undergoes translocation and mediates up-regulation of the tyrosine kinase c-Src in EGF-stimulated A431 cells."Genes to Cells. 5. 749-764 (2000)
[Publications] Sato K.-I.: "Hydrogen peroxide induces Src family tyrosine kinase-dependent egg activation process in Xenopus eggs."Develop.Growth Differ.. 43. 55-72 (2001)
[Publications] Tokmakov,A.: "Calcium oscillations in Xenopus egg cycling extracts."J.Cell.Biochem.. (印刷中).
[Publications] Dong,L.Q.: "Phosphorylation of protein kinase N by phosphoinositide-dependent protein kinase-1 mediates insulin signals to the actin cytoskeleton."Proc.Natl.Acad.Sci.USA. 97・10. 5089-5094 (2000)
[Publications] Nagumo,H.: "Rho kinase inhibitor HA-1077 prevents Rho-medicated myosin phosphatase inhibitation in smooth muscle cells."Am.J.Physiol.Cell Physiol.. 278. 57-65 (2000)
[Publications] Isagawa,T.: "Dual effects of PKNα and protein kinase C on phosphorylation of tau protein by glycogen synthase kinase-3β."Biochem.Biophys.Res.Commun.. 273・1. 209-212 (2000)
[Publications] Gao,Q.: "PKN binds and phosphorylates human Papillomavirus E6 oncoprotein."J.Biol.Chem.. 275・20. 14824-14830 (2000)
[Publications] Takahashi,M.: "Association of immature hypo-phosphorylated protein kinase Cε with an anchoring protein CG-NAP."J.Biol.Chem.. 275・44. 34592-34596 (2000)
[Publications] Misaki,K.: "PKN delays mitotic timing by inhibition of Cdc25C : Possible involvement of PKN in the regulation of cell division."Proc.Natl.Acad.Sci.USA. 98. 125-129 (2000)
[Publications] Taniguchi,T.: "Phosphorylation of tau is regulated by PKN."J.Biol.Chem.. (印刷中).
[Publications] Matsumoto,M.: "Inhibition of insulin-induced activation of Akt by a kinase-deficient mutant of the ε isozyme of protein kinase C."J.Biol.Chem.. (印刷中).

2000 Fiscal Year Annual Research Report

細胞内情報伝達機構網の研究

Principal Investigator

西塚 泰美 神戸大学, 名誉教授 (10025546)

Research Products

[Publications] Konishi,H.: "Tyrosine phosphorylation sites of protein kinase C δ in the H_2O_2-treated cells and the regulation of its catalytic activity in vitro."Proc.Natl.Acad.Sci.USA. (印刷中).

[Publications] Yamamoto,T.: "H_<>O_<>-induced tyrosine phosphorylation of protein kinase C δ by a mechanism independent of inhibition of protein-tyrosine phosphatase in CHO and COS-7 cells."Biochem.Biophys.Res.Commun.. 273・3. 960-966 (2000)

[Publications] Okuno,S.: "Studies on the phosphorylation of protein kinase B by Ca^<2+>/calmodulin-dependent protein kinases."J.Biochem.. 127・6. 965-970 (2000)

[Publications] Kitaura,J.: "Akt-dependent cytokine production in mast cells."J.Exp.Med.. 192・5. 729-740 (2000)

[Publications] Oka,M.: "Regulation of melanogenesis through phosphatidylinositol 3-kinase-Akt pathway in human G361 melanoma cells."J.Invest.Dermatol.. 115・4. 699-703 (2000)

[Publications] Oyasu,M: "Immunocytochemical localization of a neuron-specific thrombospondin-1-like protein, NELL2 : Light and electron microscopic studies in the rat brain."Mol.Brain Res.. 76. 151-160 (2000)

[Publications] Sumioka,K.: "Induction of 55 kDa PKN cleavage product by ischemia/reperfusion model in the rat retina."Invest.Opthal.Vis.Sci.. 41・1. 29-35 (2000)

[Publications] Sakai,N.: "Involvement of the actin cytoskeleton in the regulation of serotonin transporter (SET) activity : possible mechanism underlying SET regulation by protein kinase C."Neurochem.Int.. 36. 567-579 (2000)

[Publications] Shirai,Y.: "Phospholipase A_2 and its products are involved in the purinergic receptor-mediated translocation of protein kinase C in CHO-K1 cells."J.Cell Sci.. 113. 1335-1343 (2000)

[Publications] Akiyama,H.: "Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain."Brain Res.. 858. 172-176 (2000)

[Publications] Tanii,H.: "Involvement of GABA neurons in allylnitrile-induced dyskinesia."Brain Res.. 887. 454-459 (2000)

[Publications] Ueyama,T.: "cDNA cloning of an alternative splicing variant of protein kinase Cδ(PKCδIII), a new truncated form of PKCδ, in rats."Biochem.Biophys.Res.Commun.. 269・2. 557-563 (2000)

[Publications] Shirai,Y.: "Subtype-specific translocation of diacylglycerol kinase α and γ and its correlation with protein kinase C."J.Biol.Chem.. 275・32. 24760-24766 (2000)

[Publications] Nakagawa,N.: "ENH, containing PDZ and LIM domain, heart/skeletal muscle-specific protein, associates with cytoskeletal protein through its PDZ domain."Biochem.Biophys.Res.Commun.. 272. 505-512 (2000)

[Publications] Shibaguchi,H.: "Role of synaptophysin in exocytic release of dopamine from Xenopus oocytes injected with rat brain mRNA."Cellular and Molecular Neurobiology. 20. 401-408 (2000)

[Publications] Ohmori,S.: "Importance of PKC targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS)."J.Biol.Chem.. 275・34. 26449-26457 (2000)

[Publications] Larsen,E.C.: "Differential requirement for classic and novel PKC isoforms in respiratory burst and phagocytosis in RAW 264.7 cells."J.Immunol.. 165. 2809-2817 (2000)

[Publications] Yokogawa,T.: "Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface : use of immunostaining technique with monoclonal antibodies specific for PI 3,4-P_2."FEBS Lett.. 473. 222-226 (2000)

[Publications] Kondoh,T.: "In vivo gene transfer into periventricular region by electroporation."Neurologia medico-chirurgica.. 40. 618-623 (2000)

[Publications] Kajimoto,T.: "Subtype-specific translocation of the d subtype of protein kinase C and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells."Mol.Cell.Biol.. 21. 1769-1783 (2001)

[Publications] Nakai,M.: "Amyloid β protein activates PKC-δ and induces translocation of myristoylated alanine-rich C kinase substrate (MARCKS) in microglia."Neurochem.Int.. (印刷中).

[Publications] Mishima,K.: "Molecular mechanism for α2-adrenoceptor-mediated regulation of synoviocytes proliferation."Jpn.J.Pharmacol.,. (印刷中).

[Publications] Yonezawa,K: "Nutritional regulation of gene and protein expression."Curr.Opin.Clinic.Nutri.Metab.Care. 3. 253-254 (2000)

[Publications] Takahashi,T.: "Carboxyl-terminal region conserved among phosphoinositide-kinase-related kinases is indispensable for mTOR function in vivo and in vitro."Genes to Cells. 5. 765-775 (2000)

[Publications] Sato K.-I.: "Fertilization signalling and protein-tyrosine kinases."Comp.Biochem.Physiol.B.. 126. 129-148 (2000)

[Publications] Sato K.-I.: "Regulation of Xenopus oocytes, eggs, and embryos by protein-tyrosine phosphorylation."Curr.Top.Biochem.Res.. 2. 91-102 (2000)

[Publications] Sato K.-I.: "Tyrosine kinase-dependent activation of phospholipase Cγis required for calcium transient in Xenopus egg fertilization."Dev.Biol.. 224. 453-469 (2000)

[Publications] Sato K.-I.: "Adaptor protein Shc undergoes translocation and mediates up-regulation of the tyrosine kinase c-Src in EGF-stimulated A431 cells."Genes to Cells. 5. 749-764 (2000)

[Publications] Sato K.-I.: "Hydrogen peroxide induces Src family tyrosine kinase-dependent egg activation process in Xenopus eggs."Develop.Growth Differ.. 43. 55-72 (2001)

[Publications] Tokmakov,A.: "Calcium oscillations in Xenopus egg cycling extracts."J.Cell.Biochem.. (印刷中).

[Publications] Dong,L.Q.: "Phosphorylation of protein kinase N by phosphoinositide-dependent protein kinase-1 mediates insulin signals to the actin cytoskeleton."Proc.Natl.Acad.Sci.USA. 97・10. 5089-5094 (2000)

[Publications] Nagumo,H.: "Rho kinase inhibitor HA-1077 prevents Rho-medicated myosin phosphatase inhibitation in smooth muscle cells."Am.J.Physiol.Cell Physiol.. 278. 57-65 (2000)

[Publications] Isagawa,T.: "Dual effects of PKNα and protein kinase C on phosphorylation of tau protein by glycogen synthase kinase-3β."Biochem.Biophys.Res.Commun.. 273・1. 209-212 (2000)

[Publications] Gao,Q.: "PKN binds and phosphorylates human Papillomavirus E6 oncoprotein."J.Biol.Chem.. 275・20. 14824-14830 (2000)

[Publications] Takahashi,M.: "Association of immature hypo-phosphorylated protein kinase Cε with an anchoring protein CG-NAP."J.Biol.Chem.. 275・44. 34592-34596 (2000)

[Publications] Misaki,K.: "PKN delays mitotic timing by inhibition of Cdc25C : Possible involvement of PKN in the regulation of cell division."Proc.Natl.Acad.Sci.USA. 98. 125-129 (2000)

[Publications] Taniguchi,T.: "Phosphorylation of tau is regulated by PKN."J.Biol.Chem.. (印刷中).

[Publications] Matsumoto,M.: "Inhibition of insulin-induced activation of Akt by a kinase-deficient mutant of the ε isozyme of protein kinase C."J.Biol.Chem.. (印刷中).

西塚泰美神戸大学, 名誉教授 (10025546)