1997 年度実績報告書

ホスホリパーゼD(PLD)の活性制御機構と細胞活動における役割

研究課題

研究課題/領域番号	08670143
研究機関	岐阜大学
研究代表者	中島茂岐阜大学, 医学部, 助教授 (60188935)
研究分担者	坂野喜子岐阜大学, 医学部, 講師 (50116852)
キーワード	ホスホリパーゼD / プロテインキナーゼC / Rho A / アポトーシス / 細胞分化 / チロシンキナーゼ / 増殖因子 / 転写因子
研究概要	新規情報変換酵素ホスホリパーゼD(PLD)は分泌応答などの迅速応答から、細胞の増殖・分化といった中長期的応答にまで関与していることが示唆され多大な注目を集めているが、精製の困難さからその性状の解析は遅れていた。1995年末にFrohman博士ら(ニューヨーク州立大)により、最初の哺乳動物PLDとしてヒトPLD(hPLD1)遺伝子がクローニングされ、最近我々はHL60細胞のhPLD1遺伝子を解析する過程でalternative splicingを世界に先駆けて見いだし、また類似した別の遺伝子(hPLD2)の存在にも気がついた。ラット褐色細胞腫PC12細胞でも同様に2種以上の遺伝子が存在することを確認し、PC12細胞ライブラリーからrPLD1、rPLD2をクローニングし、FISH法で遺伝子座を決定した。また、HL60細胞、PC12細胞、ラットグリオーマC6細胞で分化誘導あるいはアポトーシス誘導時にPLDmRNAの発現パターンが変動することを明らかにした。一方、ブタノールやエタノールあるいはプロプラノロールといったPLDからホスファチジン酸、ジアシルグリセロール産生を抑制する試薬により、増殖因子刺激時の転写因子c-fos、c-junの発現が阻害されることから、PLDは増殖因子によるシグナル伝達経路に深く関与していることを明らかにした。一方、PLDの活性化メカニズムについても検討を加え、分化HL60細胞では走化性因子によるPLD活性化にホスファチジルイノシトール3キナーゼが関与していることを示した。また、PC12細胞では過酸化水素(H_2O_2)がチロシンリン酸化を介してPLDを活性化することを示した。今後は、さまざまなPLD変異体を発現した培養細胞を作成し、細胞増殖・癌化、分化に対するPLDの役割を明らかにする予定である。

研究成果

(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] S.Yoshimura et al.: "Differential mRNA expression of phospholipase D (PLD) isozymes during cAMP-induced differentiation in C6 glioma cells." Biochem.Biophys.Res.Commun.225. 494-499 (1996)
[文献書誌] Y.Banno et al.: "Membrane-associated phospholipase D activity in neural cell line PC12" J.Lipid Mediators Cell Signalling. 14. 237-243 (1996)
[文献書誌] K.Ojio et al.: "Effect of Clostridium difficile toxin B on IgE receptor-mediated signal transduction in rat basophilic leukemia cells:inhibition of phospholipase D activation." Biochem.Biophys.Res.Commun.224. 591-596 (1996)
[文献書誌] B.Liu et al.: "PLD activation in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA." Prostaglandins. 51. 233-248 (1996)
[文献書誌] Y.Nakamura et al.: "Brefeldin A inhibits antigen-or calcium ionophore-stimulated but not PMA-induced phospholipase D activation in rat basophilic leukemia (RBL-2H3) cells." Immunobiology. 195. 231-242 (1996)
[文献書誌] T.Kumada et al.: "Phenylarsine oxide (PAO) -mediated activation of phospholipase D in rat basophilic leukemia (RBL-2H3) cells:Possible involvement of calcium and protein kinase C." Immunobiology. 195. 347-359 (1996)
[文献書誌] T.Adachi et al.: "Phospholipase D activation in hepatocyte growth factor-stimulated rat hepatocytes;Implication in the expressions of c-jun and c-fos." Hepatology. 24. 1274-1281 (1996)
[文献書誌] K.Ohguchi et al.: "Effect of Clostridium difficile Toxin A and Toxin B on phospholipase D activation in human promyelocytic leukemic HL60 cells." Infect.Immun.64. 4433-4437 (1996)
[文献書誌] S.Nakashima et al.: "Differential expression of protein kinase C isozymes and small GTP-binding proteins during HL60 cells differentiation by retinoic acid and cyclic AMP." Immunobiology. 196. 588-598 (1996)
[文献書誌] K.Ohguchi et al.: "Enhancement of phospholipase D activity during differentiation in human promyelocytic leukemic HL60 cells:involvement of RhoA translocation to membranes." J.Biol.Chem.272. 1990-1996 (1997)
[文献書誌] Y.Ito et al.: "Implication of Ca2+-dependent protein tyrosinephosphorylation in carbachol-induced phospholipase D activation in rat pheochromocytoma PC12 cells." J.Neurochem.68. 419-425 (1997)
[文献書誌] Y.Yoshimura et al.: "Differential expression of rho family GTP-binding proteins and protein kinase C isozymes during C6 glial cell differentiation." Mol.Brain Res.45. 90-98 (1997)
[文献書誌] S.M Hammond et al.: "Characterization of two alternately spliced forms of phospholipase D1." J.Biol.Chem.272. 3860-3868 (1997)
[文献書誌] H.Sakai et al.: "Identification of differentially expressed mRNAs during rat C6 glial cell differentiation by mRNA fingerprinting using arbitrarily primed PCR (RAP)." Neurosci.Lett.229. 93-96 (1997)
[文献書誌] S.Yoshimura et al.: "Changes in the activity and mRNA levels of phospholipase D during ceramide-induced apoptosis in rat glial cells." J.Neurochem.69. 713-720 (1997)
[文献書誌] Y.Ito et al.: "Hydrogen peroxide-induced phospholipase D activation in rat pheochromocytoma PC12 cells:Possible involvement of Ca^<2+>-dependent protein tyrosine kinase." J.Neurochem.69. 729-736 (1997)
[文献書誌] M.Nakamura et al.: "Effect of wortmannin and LY294002 on N-formyl-methionyl-leucyl-phenylalanine-induced phospholipase D activation in differentiated HL60 cells." Biochem.Pharmacol.53. 1929-1936 (1997)
[文献書誌] T.Adachi et al.: "Possible involvement of pertussis toxin-sensitive G protein in hepatocyte gtowth factor-induced signal transduction in cultured rat hepatocytes." Hepatology. 26. 295-300 (1997)
[文献書誌] B.Liu et al.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing PAF receptor lacking cytoplasmic C-terminal tail." Biochem.J.327. 239-244 (1997)
[文献書誌] S.Nakashima et al.: "Molecular cloning and chromosome mapping of rat phospholipase D genes rPld1a,rPld1b and rPld2." Cytogenet.Cell Genet.(印刷中). (1998)
[文献書誌] S.Nakashima et al.: "Increased mRNA expression of phospholipase D (PLD) isozymes during granulocytic differentiation of HL60 cells." Biochim.Biophys.Acta. (印刷中). (1998)
[文献書誌] H.Sakai et al.: "mRNA fingerprinting analysis using arbitrarily primed PCR (RAP) of genes expressed during rat C6 glioma cell differentiation." Brain Tumor Pathol.(印刷中). (1998)
[文献書誌] H.Sakai et al.: "Molecular cloning of a cDNA encoding a serine protease homologous to complement C1s precursor from rat C6 glial cells and its expression during glial differentiation." Gene. (印刷中). (1998)
[文献書誌] S.Yoshimura et al.: "Ceramide formation leads to caspase-3 activation during hypoxic PC12 cell death:Inhibitory effects of Bcl-2 on ceramide formation and caspase-3 activation." J.Biol.Chem.(印刷中). (1998)

1997 年度 実績報告書

ホスホリパーゼD(PLD)の活性制御機構と細胞活動における役割

研究代表者

中島 茂 岐阜大学, 医学部, 助教授 (60188935)

研究成果

[文献書誌] S.Yoshimura et al.: "Differential mRNA expression of phospholipase D (PLD) isozymes during cAMP-induced differentiation in C6 glioma cells." Biochem.Biophys.Res.Commun.225. 494-499 (1996)

[文献書誌] Y.Banno et al.: "Membrane-associated phospholipase D activity in neural cell line PC12" J.Lipid Mediators Cell Signalling. 14. 237-243 (1996)

[文献書誌] K.Ojio et al.: "Effect of Clostridium difficile toxin B on IgE receptor-mediated signal transduction in rat basophilic leukemia cells:inhibition of phospholipase D activation." Biochem.Biophys.Res.Commun.224. 591-596 (1996)

[文献書誌] B.Liu et al.: "PLD activation in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA." Prostaglandins. 51. 233-248 (1996)

[文献書誌] Y.Nakamura et al.: "Brefeldin A inhibits antigen-or calcium ionophore-stimulated but not PMA-induced phospholipase D activation in rat basophilic leukemia (RBL-2H3) cells." Immunobiology. 195. 231-242 (1996)

[文献書誌] T.Kumada et al.: "Phenylarsine oxide (PAO) -mediated activation of phospholipase D in rat basophilic leukemia (RBL-2H3) cells:Possible involvement of calcium and protein kinase C." Immunobiology. 195. 347-359 (1996)

[文献書誌] T.Adachi et al.: "Phospholipase D activation in hepatocyte growth factor-stimulated rat hepatocytes;Implication in the expressions of c-jun and c-fos." Hepatology. 24. 1274-1281 (1996)

[文献書誌] K.Ohguchi et al.: "Effect of Clostridium difficile Toxin A and Toxin B on phospholipase D activation in human promyelocytic leukemic HL60 cells." Infect.Immun.64. 4433-4437 (1996)

[文献書誌] S.Nakashima et al.: "Differential expression of protein kinase C isozymes and small GTP-binding proteins during HL60 cells differentiation by retinoic acid and cyclic AMP." Immunobiology. 196. 588-598 (1996)

[文献書誌] K.Ohguchi et al.: "Enhancement of phospholipase D activity during differentiation in human promyelocytic leukemic HL60 cells:involvement of RhoA translocation to membranes." J.Biol.Chem.272. 1990-1996 (1997)

[文献書誌] Y.Ito et al.: "Implication of Ca2+-dependent protein tyrosinephosphorylation in carbachol-induced phospholipase D activation in rat pheochromocytoma PC12 cells." J.Neurochem.68. 419-425 (1997)

[文献書誌] Y.Yoshimura et al.: "Differential expression of rho family GTP-binding proteins and protein kinase C isozymes during C6 glial cell differentiation." Mol.Brain Res.45. 90-98 (1997)

[文献書誌] S.M Hammond et al.: "Characterization of two alternately spliced forms of phospholipase D1." J.Biol.Chem.272. 3860-3868 (1997)

[文献書誌] H.Sakai et al.: "Identification of differentially expressed mRNAs during rat C6 glial cell differentiation by mRNA fingerprinting using arbitrarily primed PCR (RAP)." Neurosci.Lett.229. 93-96 (1997)

[文献書誌] S.Yoshimura et al.: "Changes in the activity and mRNA levels of phospholipase D during ceramide-induced apoptosis in rat glial cells." J.Neurochem.69. 713-720 (1997)

[文献書誌] Y.Ito et al.: "Hydrogen peroxide-induced phospholipase D activation in rat pheochromocytoma PC12 cells:Possible involvement of Ca^<2+>-dependent protein tyrosine kinase." J.Neurochem.69. 729-736 (1997)

[文献書誌] M.Nakamura et al.: "Effect of wortmannin and LY294002 on N-formyl-methionyl-leucyl-phenylalanine-induced phospholipase D activation in differentiated HL60 cells." Biochem.Pharmacol.53. 1929-1936 (1997)

[文献書誌] T.Adachi et al.: "Possible involvement of pertussis toxin-sensitive G protein in hepatocyte gtowth factor-induced signal transduction in cultured rat hepatocytes." Hepatology. 26. 295-300 (1997)

[文献書誌] B.Liu et al.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing PAF receptor lacking cytoplasmic C-terminal tail." Biochem.J.327. 239-244 (1997)

[文献書誌] S.Nakashima et al.: "Molecular cloning and chromosome mapping of rat phospholipase D genes rPld1a,rPld1b and rPld2." Cytogenet.Cell Genet.(印刷中). (1998)

[文献書誌] S.Nakashima et al.: "Increased mRNA expression of phospholipase D (PLD) isozymes during granulocytic differentiation of HL60 cells." Biochim.Biophys.Acta. (印刷中). (1998)

[文献書誌] H.Sakai et al.: "mRNA fingerprinting analysis using arbitrarily primed PCR (RAP) of genes expressed during rat C6 glioma cell differentiation." Brain Tumor Pathol.(印刷中). (1998)

[文献書誌] H.Sakai et al.: "Molecular cloning of a cDNA encoding a serine protease homologous to complement C1s precursor from rat C6 glial cells and its expression during glial differentiation." Gene. (印刷中). (1998)

[文献書誌] S.Yoshimura et al.: "Ceramide formation leads to caspase-3 activation during hypoxic PC12 cell death:Inhibitory effects of Bcl-2 on ceramide formation and caspase-3 activation." J.Biol.Chem.(印刷中). (1998)

1997 年度実績報告書

中島茂岐阜大学, 医学部, 助教授 (60188935)