1996 Fiscal Year Annual Research Report

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

Research Project

Project/Area Number	08670143
Research Category	Grant-in-Aid for Scientific Research (C)
Research Institution	Gifu University
Principal Investigator	中島茂岐阜大学, 医学部, 講師 (60188935)
Co-Investigator(Kenkyū-buntansha)	坂野喜子岐阜大学, 医学部, 助手 (50116852)
Keywords	ホスホリパーゼD / プロテインキナーゼC / RhoA / クロストリジウムトキシンB / 細胞分化 / チロシンキナーゼ
Research Abstract	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た。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(hPLD1に相当するrPLD1にはやはりalternative splicingがみられる)、現在PC12細胞ライブラリーからrPLD1、rPLD2をクローニングをほぼ終えた。また、degenerate primersを用いたRT-PCR法により新規PLD遺伝子の探索を行っている。HL60細胞、PC12細胞、ラットグリオーマC6細胞で分化誘導によりPLD mRNAの発現パターンが変動するという知見を得ており、現在発現パターンの確認とともに、PLD活性の変化を検討中である。一方、手術により摘出されたヒトガン組織や種々の培養細胞でPLD mRNAの発現パターンについても検討している。まだ結論するには時期尚早であるが、腺ガン、扁平上皮ガンなど組織型により、また細胞の悪性度により発現パターンに一定の傾向がみられる。また、悪性度の高い脳腫瘍グラオブラストーマの細胞株ではhPLD1に点突然変異があることを見出し、摘出組織での変異を現在検討している。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。

Research Products

(14 results)

All Other

All Publications (14 results)

[Publications] 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)
[Publications] Y.Banno et al: "Membrane-associated phospholipase D activity in neural cell line PC12." J.Lipid Mediators Cell Signalling. 14. 237-243 (1996)
[Publications] 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)
[Publications] B.Liu et al.: "PLD activation in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA." Prostaglandins. 51. 233-248 (1996)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] S.Nakashima et al.: "Differential expression of protein kinase C isozymes and small GTP-binding proteins during HL-60 cells differentiation by retinoic acid and cyclic AMP." Immunobiology. 196. 588-598 (1996)
[Publications] K.Ohguchi et al.: "Enhancement to phospholipase D activity during differentiation in human promyelocytic leukemic HL60 cells : involvement of RhoA translocation to membranes." J.Biol.Chem.272. 1990-1996 (1997)
[Publications] 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)
[Publications] 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.(印刷中). (1997)
[Publications] S.M.Hammond et al.: "Characterization of two alternately spliced forms of phospholipase D1." J.Biol.Chem.(印刷中). (1997)
[Publications] 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.(印刷中). (1997)

1996 Fiscal Year Annual Research Report

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

Principal Investigator

中島 茂 岐阜大学, 医学部, 講師 (60188935)

Research Products

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] S.Nakashima et al.: "Differential expression of protein kinase C isozymes and small GTP-binding proteins during HL-60 cells differentiation by retinoic acid and cyclic AMP." Immunobiology. 196. 588-598 (1996)

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

[Publications] 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)

[Publications] 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.(印刷中). (1997)

[Publications] S.M.Hammond et al.: "Characterization of two alternately spliced forms of phospholipase D1." J.Biol.Chem.(印刷中). (1997)

[Publications] 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.(印刷中). (1997)

中島茂岐阜大学, 医学部, 講師 (60188935)