1993 Fiscal Year Annual Research Report

発癌過程ならびに癌細胞形質の発現におけるプロテインホスファターゼの意義

Research Project

Project/Area Number	05151004
Research Institution	Hokkaido University
Principal Investigator	菊池九二三北海道大学, 免疫科学研究所, 教授 (20006117)
Co-Investigator(Kenkyū-buntansha)	日野田裕治札幌医科大学, 医学部, 講師 (10165128) 菅沼雅美埼玉県立がんセンター, 研究所, 研究員 (20196695) 島礼国立がんセンター, 研究所, 室長 (10196462) 田村眞理東北大学, 加齢医学研究所, 教授 (20124604) 武田誠郎広島大学, 医学部, 教授 (40030853)
Keywords	プロテインホスファターゼ / セリン / スレオニンホスファターゼ / チロシンホスファターゼ / 発癌 / 癌形質 / 細胞増殖 / 蛋白脱リン後 / 遺伝子発現
Research Abstract	菊池らは,セリン/スレオニン残基特異的プロテインホスファターゼPP1について次の点を明らかにした。 1)腹水肝癌細胞でPP1のイソホームPP1αのmRNAが特異的に上昇する。2)腹水肝癌細胞でPP1αの酵素蛋白が,検索した細胞株すべてにおいて例外なく高発現していた。3)再生肝では,術後12時間のG1/S移行期で,核内PP1活性が特異的かつ一過性に上昇する。4)EGF刺激後の初代培養肝細胞でも,EGF刺激後40時間のG1/S移行期に一過性にPP1活性の特異的上昇を観察した。5)この核内活性上昇の機序は,翻訳段階での亢進と核内移行後の2次修飾による活性化の2つが考えられる。6)IL-2によるT細胞刺激では,刺激後5,10,15分とPP1型活性の低下がみられた。このPP1活性の減少は可逆的かつ一過性で,可溶画分についてのみ見られ,顆粒画分では不変であった。またPP2A,PP2Cは不変できわめて特異的であった。以上より本酵素の癌性変異の特徴は,特定の分子種PP1αについて転写レベルの亢進によるもので,その結果として翻訳段階でも上昇が認められた。この点,正常組織である再生肝やIL-2による増殖刺激でみられる応答と機構を異にする。菅沼は本酵素阻害剤ノデュラリンやマイクロシスチンが,junやfosなどの初期応答遺伝子を誘導することを初代培養肝細胞で見いだした。武田はPP2Aの新しいホロ酵素を分離精製した。田村はPP2CβのcDNAの塩基配列を決定した。島はPP2Aの制御サブユニットのクローニングに成功し,腫瘍での発現を調べ,久野は制御サブユニットのクローニングを行った。チロシンホスファターゼでは,野口はLRPのアンチセンスDNAを導入して脱癌を観察し,日野田はPTPG1により逆に悪性形質の増大を観察し,分子種により発癌・制癌の両方向に作用し得る。矢倉は分化過程で異なる効果を観察した。伊藤はミオシン結合型PP1の組織化学,小林はPP2Cのリン酸化部位を推定した。

Research Products

(32 results)

All Other

All Publications (32 results)

[Publications] Norio Takizawa: "Selective increases in isoform PP1α of type-1 protein phosphatase in ascites hepatoma cells." Japanese Journal of Cancer Research. 85(印刷中). (1994)
[Publications] Mostafa Saasat: "Gene expression of protein phosphatases in rat ascites hepatoma cell lines." Cancer Detection and Prevention. 18(印刷中). (1994)
[Publications] Chika Katagiri: "Increases in protein phosphatase 2B activity in lymphoid tissues and T-lymphocytes of autoimmune MRL/MpJ-lpr/lpr mice." Journal ofBiochemistry. 114. 874-878 (1993)
[Publications] Kunimi Kikuchi: "Expressions and activities of protein phosphatases during carcinogenesis,regeneration,and cell cycle." Advances in Protein Phosphatases. 7. 221-236 (1993)
[Publications] Shu-ichi Matsuzawa: "IL-2 induces transient and specific decrease in cytosolic protein phosphatase PP1 activity in murine T cell lines." Biochemical and Biophysical Research Communications. 195. 989-996 (1993)
[Publications] Tang Zhu: "Alterations in activities of protein phosphatases PP1 and PP2A in T and B lymphocytes of autoimmune MRL/MpJ-lpr/lpr mice." Journal of Biochemistry. 114. 50-54 (1993)
[Publications] T.Murakami: "Characterization of multiple molecular forms of Mg^<2+>-dependent protein phosphatase from Saccharomyces cerevisiae." J. Biochem.(印刷中).
[Publications] T.Terasawa: "Molecular cloning of a novel isotype of Mg^<2+>-dependent protein phosphatase β (type 2Cβ) enriched in brain and heart." Arch.Biochem.Biophys.307. 342-349 (1993)
[Publications] K.Fukunaga: "Deposphorylation of autophosphorylated Ca^<2+>/calmodulin-dependent protein kinase II by protein phosphatase 2C." J.Biol.Chem.268. 133-137 (1993)
[Publications] C.D.Chang: "cDNA cloning of an alternatively spliced isoform of the regulatory subunit of Ca^<2+>/calmodulin-dependent protin phosphatase (calcineurin Bα2)." Biochim.Biophys.Acta. (印刷中).
[Publications] H.Mukai: "FKBP12-FK506 complex inhibits phosphatase activity of two mammalian isoforms of calcineurin irrespective of their substrates or activation mechanisms." J.Biochem.113. 292-298 (1993)
[Publications] M.Asami: "Detection of the FK506-FKBP-calcineurin complex by a simple binding assay." Biochem.Biophys.Res.Commun.192. 1388-1394 (1993)
[Publications] J.Yatsunami: "Hyperphosphorylation of retinoblastoma protin and p53 by okadaic acid,a tumor promoter." Cancer Res.53. 239-241 (1993)
[Publications] Masami Suganuma: "Absence of synergistic effects on tumor promotion in CD-1 mouse skin by simultaneous applications of two different types of tumor promoters,okadaic acid and teleocidin." Cancer Res.53. 1012-1016 (1993)
[Publications] Hirota Fujiki: "Tumor promotion by inhibitors of protein phosphatases 1 and 2A,the okadaic acid class of compounds." Adv.Cancer Res.61. 143-194 (1993)
[Publications] Y.S.Chun: "PP1γ2,a testis specific serine/threonine protein phosphatase type 1 catalytic subunit,is associated with GRP 78,a member of hsp 70 family." Proc.Natl.Acad.Sci.USA. 印刷中.
[Publications] Hiroshi Shima: "Protein phosphatase 1γ2 is associated with nuclei of meiotic cells in rat testis." Biochem.Biophys.Res.Commun.194. 930-937 (1993)
[Publications] Hiroshi Shima: "Identification of PP1 catalytic isotypes PP1γ1,PP1δ and PP1α in various rat tissues." Biochem.Biophys.Res.Commun.192. 1289-1296 (1993)
[Publications] M.Ogimoto: "Negative regulation of apoptotic death in immature B cells by CD45." Int.Immunol.(印刷中).
[Publications] K.Hasegawa: "Chromosomal assignment of the gene for protein tyrosine phosphatase HPTPδ." Jpn.J.Cancer Res.84. 1219-1222 (1993)
[Publications] M.Ogimoto: "Induction of CD45 isoform switch in murine B cells by antigen receptor stimulation and by phorbol myristate acetate and ionomycin." Cell.Immunol.151. 97-109 (1993)
[Publications] Z.Wang: "Transcriptional regulatory regions for expression of the rat pyruvate kinase M gene." Eur.J.Biochem.(印刷中).
[Publications] Tamio Noguchi: "The regulation of gene expression by insulin is differentially impaired in the liver of the genetically obese-hyperglycemic Wistar fatty rat." FEBS Lett.328. 145-148 (1993)
[Publications] Tamio Noguchi: "An enhancer unit of L-type pyruvate kinase gene is responsible for transcriptional stimulation by dietary fructose as well as glucose in transgenic mice." FEBS Lett.318. 269-272 (1993)
[Publications] Kazuhito Ichikawa: "Calponin phosphatase in smooth muscle: a possible role of type 1 protein phosphatase in smooth muscle relaxation." Biochem.Biophys.Res.Commun.193. 827-833 (1993)
[Publications] Hiroo Itoh: "Inhibition of myosin light chain phosphatase during Ca^<2+>-independent vasocontraction." Am.J.Physiol.265. C1319-C1324 (1993)
[Publications] Satoshi Kanoh: "Actin-binding peptide from smooth muscle myosin light chain kinase." Biochemistry. 32. 8902-8907 (1993)
[Publications] M.Takekawa: "Chromosomal localization of protein tyrosine phosphatase G1 gene and characterization of the abberant transcripts in human colon cancer cells." FEBS Lett.(印刷中). (1994)
[Publications] M.Adachi: "Induction of protein-tyrosine phosphatase LC-PTP by IL-2 in human T cells.LC-PTP is an early response gene." FEBS Lett.338. 47-52 (1994)
[Publications] F.Itoh: "Expression and chromosomal assignment of PTPH1 gene encoding a cytosolic protein tyrosine phosphatase homologous to cytoskeletal-associaated proteins." Int.J.Cancer. 55. 947-951 (1993)
[Publications] Takayasu Kobayashi: "Expression of rat type 2Cα protein phosphatase in Saccharomyces cerevisiae cells." Adv.Prot.Phosphatases. 7. 477-487 (1993)
[Publications] Takayasu Kobayashi: "Phophorylation of Mg^<2+>-dependent protein phosphatase α (type 2Cα) by casein kinase II." Biochem.Biophys.Res.Commun.195. 484-489 (1993)

1993 Fiscal Year Annual Research Report

発癌過程ならびに癌細胞形質の発現におけるプロテインホスファターゼの意義

Principal Investigator

菊池 九二三 北海道大学, 免疫科学研究所, 教授 (20006117)

Research Products

[Publications] Norio Takizawa: "Selective increases in isoform PP1α of type-1 protein phosphatase in ascites hepatoma cells." Japanese Journal of Cancer Research. 85(印刷中). (1994)

[Publications] Mostafa Saasat: "Gene expression of protein phosphatases in rat ascites hepatoma cell lines." Cancer Detection and Prevention. 18(印刷中). (1994)

[Publications] Chika Katagiri: "Increases in protein phosphatase 2B activity in lymphoid tissues and T-lymphocytes of autoimmune MRL/MpJ-lpr/lpr mice." Journal ofBiochemistry. 114. 874-878 (1993)

[Publications] Kunimi Kikuchi: "Expressions and activities of protein phosphatases during carcinogenesis,regeneration,and cell cycle." Advances in Protein Phosphatases. 7. 221-236 (1993)

[Publications] Shu-ichi Matsuzawa: "IL-2 induces transient and specific decrease in cytosolic protein phosphatase PP1 activity in murine T cell lines." Biochemical and Biophysical Research Communications. 195. 989-996 (1993)

[Publications] Tang Zhu: "Alterations in activities of protein phosphatases PP1 and PP2A in T and B lymphocytes of autoimmune MRL/MpJ-lpr/lpr mice." Journal of Biochemistry. 114. 50-54 (1993)

[Publications] T.Murakami: "Characterization of multiple molecular forms of Mg^<2+>-dependent protein phosphatase from Saccharomyces cerevisiae." J. Biochem.(印刷中).

[Publications] T.Terasawa: "Molecular cloning of a novel isotype of Mg^<2+>-dependent protein phosphatase β (type 2Cβ) enriched in brain and heart." Arch.Biochem.Biophys.307. 342-349 (1993)

[Publications] K.Fukunaga: "Deposphorylation of autophosphorylated Ca^<2+>/calmodulin-dependent protein kinase II by protein phosphatase 2C." J.Biol.Chem.268. 133-137 (1993)

[Publications] C.D.Chang: "cDNA cloning of an alternatively spliced isoform of the regulatory subunit of Ca^<2+>/calmodulin-dependent protin phosphatase (calcineurin Bα2)." Biochim.Biophys.Acta. (印刷中).

[Publications] H.Mukai: "FKBP12-FK506 complex inhibits phosphatase activity of two mammalian isoforms of calcineurin irrespective of their substrates or activation mechanisms." J.Biochem.113. 292-298 (1993)

[Publications] M.Asami: "Detection of the FK506-FKBP-calcineurin complex by a simple binding assay." Biochem.Biophys.Res.Commun.192. 1388-1394 (1993)

[Publications] J.Yatsunami: "Hyperphosphorylation of retinoblastoma protin and p53 by okadaic acid,a tumor promoter." Cancer Res.53. 239-241 (1993)

[Publications] Masami Suganuma: "Absence of synergistic effects on tumor promotion in CD-1 mouse skin by simultaneous applications of two different types of tumor promoters,okadaic acid and teleocidin." Cancer Res.53. 1012-1016 (1993)

[Publications] Hirota Fujiki: "Tumor promotion by inhibitors of protein phosphatases 1 and 2A,the okadaic acid class of compounds." Adv.Cancer Res.61. 143-194 (1993)

[Publications] Y.S.Chun: "PP1γ2,a testis specific serine/threonine protein phosphatase type 1 catalytic subunit,is associated with GRP 78,a member of hsp 70 family." Proc.Natl.Acad.Sci.USA. 印刷中.

[Publications] Hiroshi Shima: "Protein phosphatase 1γ2 is associated with nuclei of meiotic cells in rat testis." Biochem.Biophys.Res.Commun.194. 930-937 (1993)

[Publications] Hiroshi Shima: "Identification of PP1 catalytic isotypes PP1γ1,PP1δ and PP1α in various rat tissues." Biochem.Biophys.Res.Commun.192. 1289-1296 (1993)

[Publications] M.Ogimoto: "Negative regulation of apoptotic death in immature B cells by CD45." Int.Immunol.(印刷中).

[Publications] K.Hasegawa: "Chromosomal assignment of the gene for protein tyrosine phosphatase HPTPδ." Jpn.J.Cancer Res.84. 1219-1222 (1993)

[Publications] M.Ogimoto: "Induction of CD45 isoform switch in murine B cells by antigen receptor stimulation and by phorbol myristate acetate and ionomycin." Cell.Immunol.151. 97-109 (1993)

[Publications] Z.Wang: "Transcriptional regulatory regions for expression of the rat pyruvate kinase M gene." Eur.J.Biochem.(印刷中).

[Publications] Tamio Noguchi: "The regulation of gene expression by insulin is differentially impaired in the liver of the genetically obese-hyperglycemic Wistar fatty rat." FEBS Lett.328. 145-148 (1993)

[Publications] Tamio Noguchi: "An enhancer unit of L-type pyruvate kinase gene is responsible for transcriptional stimulation by dietary fructose as well as glucose in transgenic mice." FEBS Lett.318. 269-272 (1993)

[Publications] Kazuhito Ichikawa: "Calponin phosphatase in smooth muscle: a possible role of type 1 protein phosphatase in smooth muscle relaxation." Biochem.Biophys.Res.Commun.193. 827-833 (1993)

[Publications] Hiroo Itoh: "Inhibition of myosin light chain phosphatase during Ca^<2+>-independent vasocontraction." Am.J.Physiol.265. C1319-C1324 (1993)

[Publications] Satoshi Kanoh: "Actin-binding peptide from smooth muscle myosin light chain kinase." Biochemistry. 32. 8902-8907 (1993)

[Publications] M.Takekawa: "Chromosomal localization of protein tyrosine phosphatase G1 gene and characterization of the abberant transcripts in human colon cancer cells." FEBS Lett.(印刷中). (1994)

[Publications] M.Adachi: "Induction of protein-tyrosine phosphatase LC-PTP by IL-2 in human T cells.LC-PTP is an early response gene." FEBS Lett.338. 47-52 (1994)

[Publications] F.Itoh: "Expression and chromosomal assignment of PTPH1 gene encoding a cytosolic protein tyrosine phosphatase homologous to cytoskeletal-associaated proteins." Int.J.Cancer. 55. 947-951 (1993)

[Publications] Takayasu Kobayashi: "Expression of rat type 2Cα protein phosphatase in Saccharomyces cerevisiae cells." Adv.Prot.Phosphatases. 7. 477-487 (1993)

[Publications] Takayasu Kobayashi: "Phophorylation of Mg^<2+>-dependent protein phosphatase α (type 2Cα) by casein kinase II." Biochem.Biophys.Res.Commun.195. 484-489 (1993)

菊池九二三北海道大学, 免疫科学研究所, 教授 (20006117)