1990 年度実績報告書

造血因子の構造と作用の分子生物学的検討と生体内作用

研究課題

研究課題/領域番号	02256101
研究機関	東京大学
研究代表者	新井賢一東京大学, 医科学研究所, 教授 (00012782)
研究分担者	高津聖志熊本大学, 医学部, 教授 (10107055) 長田重一 (財)大阪バイオサイエンス研究所, 部長 (70114428) 平野俊夫大阪大学, 医学部, 教授 (40136718) 高久史麿東京大学, 医学部, 教授 (40048955)
キーワード	造血因子 / インタ-ロイキン / コロニ-刺激因子 / レセプタ- / 情報伝達 / 発現クロ-ニング / 分子生物学 / サイトカイン
研究概要	ILー3,GMーCSFレセプタ-は高親和性、低親和性の2種類からなり、その活性にはチロシンキナ-ゼが関与している。マウス低親和性ILー3レセプタ-cDNAを単離し、この分子にはチロシンキナ-ゼドメインが存在しないことを示した。ヒト高親和性GMーCSFレセプタ-は少なくともα,β鎖からなるが、我々は低親和性マウスILー3レセプタ-と高いホモロジ-を持つβ鎖cDNAクロ-ンの単離に成功し、α鎖cDNAクロ-ンとの共発現により高親和性レセプタ-の再構成を行なった。α鎖は血液細胞のみならず胎盤や各種固形癌細胞株に豊富に発現している。α鎖のみを発現している細胞ではILー3はGMーCSFと全く競合しないが、α鎖とβ鎖を発現する細胞においてはILー3によってGMーCSFの結合が部分的に阻害され、ヒトの系ではILー3とGMーCSFレセプタ-がリガンド結合ユニットを共有することを示した。 ILー6レセプタ-は80kDのILー6結合分子とgp130のシグナル伝達分子より構成されていることを示した。単クロ-ン抗体を用いてヒトgp130cDNAを単離し、896アミノ酸より成るこの分子の細胞内領域が277アミノ酸より成ることを示した。gp130分子そのものはILー6結合能はないが80kD分子と共に高親和性結合部位を形成する。発現クロ-ニング法によりGーCSFレセプタ-cDNAの単離に成功しマウスは812、ヒトは813アミノ酸からなることを示した。その細胞外領域にはサイトカインレセプタ-ファミリ-に共通する約200アミノ酸残基の領域、3個のフィブロネクチンtypeIII領域が存在した。 ILー5依存性プレB細胞株(T88ーM)より得た抗ILー5レセプタ-抗体H7を用いた発現クロ-ニング法によりマウスILー5レセプタ-cDNAの単離を行なった。このcDNAを導入されたCOS7細胞は低親和性ILー5レセプタ-を発現したが、ILー3依存性株化細胞FDCーP1は高親和性、低親和性の2種類のILー5レセプタ-を発現し、ILー5に応答して増殖した。

研究成果

(31件)

すべてその他

すべて文献書誌 (31件)

[文献書誌] Nishida,J.: "Deinition of GC rich motif as regulatory sequence of the human ILー3 gene:Coordinate regulation of the ILー3 gene by CLE2/GC box of the GMーCSF gene in T cell activation" Int Immunol.(1991)
[文献書誌] Hayashida,K.: "Molecular cloning of a second subunit of the human GMーCSF receptor:Resonsitution of a high affinity GMーCSF receptor by coーexpression of the low affinity GMーCSF receptor and an ILー3 receptorーlike molecule." Proc,Natl Acad Sci U S A.87. 9655-9659 (1990)
[文献書誌] Itoh,N.: "Molecular cloning of a cDNA encoding the murine interleukinー3 receptor reveals a consensus motif for a new cytokine receptor gene family." Science. 247. 324-327 (1990)
[文献書誌] Gorman,D.M.: "Cloning and expression of a gene encoding an interleukin 3 receptorーlike protein:Identification of another member of the cytokine receptor gene family." Proc Natl Acad Sci U S A.87. 5459-5463 (1990)
[文献書誌] Sugimoto,K.: "Inducible and nonーinducible factors coーoperatively active the GMーCSF promoter by interacting with two adjacent DNA motifs." Int Immunol.2. 787-794 (1990)
[文献書誌] Arai,K.: "Cytokines;Cordinators of immune and inflammatory responses." Ann Rev Biochem. 59. 783-836 (1990)
[文献書誌] Chiba,S.,<Takaku,F.>___ー and et al.: "Characterization and molecular features of the cell surface receptor for human granulocyteーmacrophage colonyーstimulating factor." Leukemia. 4. 29-36 (1990)
[文献書誌] Chiba,S.,.,<Takaku,F.>___ー and et al.: "Identification and cellular distribution of distinct proteins forming human GMーCSF receptor" Cell Regulation. 1. 327-335 (1990)
[文献書誌] Miyagawa,K.,<Takaku,F.>___ー and et al.: "Frequent expression of receptors for granulocyteーmacrophage colonyーstimulating factor on human nonhematopoietic tumor cell lines." J.Cell.Physiol.143. 483-487 (1990)
[文献書誌] Chiba,S.,<Takaku,F.>___ー and et al.: "Affinity purification of human granulocyteーmacrophage colonyーstimulating factor receptor alphaーchain:demonstration of binding by photoaffinity labeling." J.Biol.Chem.265. 19777-19781 (1990)
[文献書誌] Taketazu,F.,<Takaku,F.>___ー and et al.: "ILー3 specifically inhibits GMーCSF binding to the higher affinity receptor." J.Cell.Physiol.(1991)
[文献書誌] Shimano,H.: "Human monocyte colonyーstimulating factor enhances the clearance of lipoproteins containing apolipoprotein B100 via both low density lipoprotein receptorーdependent and independent pathways." J Biol Chem. 265. 12869-12875 (1990)
[文献書誌] Sugita,T.et al: "Functional murine ILー6 receptor with the intracisternal Aーparticle gene product at its cytoplasmic domain:Its possible role in plasmacytomagenesis." J.Exp.Med.171. 2001-2009 (1990)
[文献書誌] Yasukawa,K.et al: "Purification and Characterization of Soluble Human ILー6 Receptor Expressed in CHO Cells." J.Biochem.108. 673-676 (1990)
[文献書誌] Hibi,M.et al: "Molecular cloning and expression of an ILー6 signal transducer,gp130." Cell. 63. 1149-1157 (1990)
[文献書誌] Hirano,T.et al: "Biological and clinical aspects of interleukin 6." Immunology Today. 11. 443-449 (1990)
[文献書誌] Toyama,R.: "Human chorionic gonadotropin alpha and human cytomegalovirus promoters are extremely active in the fission yeast S.pombe." FEBS Lett. 268. 217-221 (1990)
[文献書誌] M.Nishizawa et al.: "Multiple elements in the promoter of granulocyte colonyーstimulating factor gene regulate its constitutive expression in human carcinoma cells." J.Biol.Chem.265. 5897-5902 (1990)
[文献書誌] R.Fukunaga et al.: "Expression cloning of a receptor for murine granulocyte colonyーstimulating factor." Cell. 61. 341-350 (1990)
[文献書誌] M.Nishizawa & S.Nagata: "Regulatory elements responsible for inducible expression of granulocyte colonyーstimulating factor gene in macrophages." Mol.Cell.Biol.10. 2002-2011 (1990)
[文献書誌] R.Fukunaga et al.: "Purification and characterization of the receptor for murine granulocyte colonyーstimulating factor." J.Biol.Chem.265. 14008-14015 (1990)
[文献書誌] S.Mizushima & S.Nagata: "pEFーBOS:a powerful mammalian expression vector." Nucleic Acids Res.18. 5322 (1990)
[文献書誌] R.Fukunaga et al.: "Three different mRNAs encoding human granulocyte colonyーstimulating factor receptor." Proc.Natl.Acad.Sci.USA. 87. 8702-8706 (1990)
[文献書誌] Yamaguchi,N.: "Characterization of murine ILー5 receptors by using monoclonal antiーreceptor antibody." International Immunology. 2. 181-187 (1990)
[文献書誌] Hitoshi,Y.: "Distribution of ILー5 receptorーpositive B cells:expression of ILー5 receptor on Lyー(CD5)^+ B cells." The Journal of Immunology. 144. 4218-4225 (1990)
[文献書誌] Yamaguchi,Y.: "In vitro and in vivo induction of ILー5 mRNA by ILー2 that causes eosinophilia in mice." The Journal of Immunology. 145. 873-877 (1990)
[文献書誌] Katoh,S.: "Conversion of Lyー1ーpositive Bーlineage cells into Lyー1ーpositive macrophages in longーterm bone marrow cultures." Developmental Immunology. 1. 113-125 (1990)
[文献書誌] Takaki,S.: "Molecular cloning and expression of the murine interleukinー5 receptor." The EMBO Journal. 9. 4367-4374 (1990)
[文献書誌] Murata,Y.: "Interleukin 5 and interleukin 3 induce serine and tyrosine phosphorylations of several cellular proteins in an interleukin 5 dependent cell line." Biochemical and Biophysical Recearch Communications. 173. 1102-1108 (1990)
[文献書誌] Yokota,T.: "Handbook of Experimental Pharmacology,Vol.95/I Peptide Growth.Factors and Their Receptors I" SpringerーVerlag, 31 (1990)
[文献書誌] Hirano,T.,and T.Kishimoto: "Handbook of Experimental Pharmacology vol.95/I “Peptide Growth Factors and Their Receptors"" SpringerーVerlag, 35 (1990)

1990 年度 実績報告書

造血因子の構造と作用の分子生物学的検討と生体内作用

研究代表者

新井 賢一 東京大学, 医科学研究所, 教授 (00012782)

研究成果

[文献書誌] Nishida,J.: "Deinition of GC rich motif as regulatory sequence of the human ILー3 gene:Coordinate regulation of the ILー3 gene by CLE2/GC box of the GMーCSF gene in T cell activation" Int Immunol.(1991)

[文献書誌] Hayashida,K.: "Molecular cloning of a second subunit of the human GMーCSF receptor:Resonsitution of a high affinity GMーCSF receptor by coーexpression of the low affinity GMーCSF receptor and an ILー3 receptorーlike molecule." Proc,Natl Acad Sci U S A.87. 9655-9659 (1990)

[文献書誌] Itoh,N.: "Molecular cloning of a cDNA encoding the murine interleukinー3 receptor reveals a consensus motif for a new cytokine receptor gene family." Science. 247. 324-327 (1990)

[文献書誌] Gorman,D.M.: "Cloning and expression of a gene encoding an interleukin 3 receptorーlike protein:Identification of another member of the cytokine receptor gene family." Proc Natl Acad Sci U S A.87. 5459-5463 (1990)

[文献書誌] Sugimoto,K.: "Inducible and nonーinducible factors coーoperatively active the GMーCSF promoter by interacting with two adjacent DNA motifs." Int Immunol.2. 787-794 (1990)

[文献書誌] Arai,K.: "Cytokines;Cordinators of immune and inflammatory responses." Ann Rev Biochem. 59. 783-836 (1990)

[文献書誌] Chiba,S.,<Takaku,F.>___ー and et al.: "Characterization and molecular features of the cell surface receptor for human granulocyteーmacrophage colonyーstimulating factor." Leukemia. 4. 29-36 (1990)

[文献書誌] Chiba,S.,.,<Takaku,F.>___ー and et al.: "Identification and cellular distribution of distinct proteins forming human GMーCSF receptor" Cell Regulation. 1. 327-335 (1990)

[文献書誌] Miyagawa,K.,<Takaku,F.>___ー and et al.: "Frequent expression of receptors for granulocyteーmacrophage colonyーstimulating factor on human nonhematopoietic tumor cell lines." J.Cell.Physiol.143. 483-487 (1990)

[文献書誌] Chiba,S.,<Takaku,F.>___ー and et al.: "Affinity purification of human granulocyteーmacrophage colonyーstimulating factor receptor alphaーchain:demonstration of binding by photoaffinity labeling." J.Biol.Chem.265. 19777-19781 (1990)

[文献書誌] Taketazu,F.,<Takaku,F.>___ー and et al.: "ILー3 specifically inhibits GMーCSF binding to the higher affinity receptor." J.Cell.Physiol.(1991)

[文献書誌] Shimano,H.: "Human monocyte colonyーstimulating factor enhances the clearance of lipoproteins containing apolipoprotein B100 via both low density lipoprotein receptorーdependent and independent pathways." J Biol Chem. 265. 12869-12875 (1990)

[文献書誌] Sugita,T.et al: "Functional murine ILー6 receptor with the intracisternal Aーparticle gene product at its cytoplasmic domain:Its possible role in plasmacytomagenesis." J.Exp.Med.171. 2001-2009 (1990)

[文献書誌] Yasukawa,K.et al: "Purification and Characterization of Soluble Human ILー6 Receptor Expressed in CHO Cells." J.Biochem.108. 673-676 (1990)

[文献書誌] Hibi,M.et al: "Molecular cloning and expression of an ILー6 signal transducer,gp130." Cell. 63. 1149-1157 (1990)

[文献書誌] Hirano,T.et al: "Biological and clinical aspects of interleukin 6." Immunology Today. 11. 443-449 (1990)

[文献書誌] Toyama,R.: "Human chorionic gonadotropin alpha and human cytomegalovirus promoters are extremely active in the fission yeast S.pombe." FEBS Lett. 268. 217-221 (1990)

[文献書誌] M.Nishizawa et al.: "Multiple elements in the promoter of granulocyte colonyーstimulating factor gene regulate its constitutive expression in human carcinoma cells." J.Biol.Chem.265. 5897-5902 (1990)

[文献書誌] R.Fukunaga et al.: "Expression cloning of a receptor for murine granulocyte colonyーstimulating factor." Cell. 61. 341-350 (1990)

[文献書誌] M.Nishizawa & S.Nagata: "Regulatory elements responsible for inducible expression of granulocyte colonyーstimulating factor gene in macrophages." Mol.Cell.Biol.10. 2002-2011 (1990)

[文献書誌] R.Fukunaga et al.: "Purification and characterization of the receptor for murine granulocyte colonyーstimulating factor." J.Biol.Chem.265. 14008-14015 (1990)

[文献書誌] S.Mizushima & S.Nagata: "pEFーBOS:a powerful mammalian expression vector." Nucleic Acids Res.18. 5322 (1990)

[文献書誌] R.Fukunaga et al.: "Three different mRNAs encoding human granulocyte colonyーstimulating factor receptor." Proc.Natl.Acad.Sci.USA. 87. 8702-8706 (1990)

[文献書誌] Yamaguchi,N.: "Characterization of murine ILー5 receptors by using monoclonal antiーreceptor antibody." International Immunology. 2. 181-187 (1990)

[文献書誌] Hitoshi,Y.: "Distribution of ILー5 receptorーpositive B cells:expression of ILー5 receptor on Lyー(CD5)^+ B cells." The Journal of Immunology. 144. 4218-4225 (1990)

[文献書誌] Yamaguchi,Y.: "In vitro and in vivo induction of ILー5 mRNA by ILー2 that causes eosinophilia in mice." The Journal of Immunology. 145. 873-877 (1990)

[文献書誌] Katoh,S.: "Conversion of Lyー1ーpositive Bーlineage cells into Lyー1ーpositive macrophages in longーterm bone marrow cultures." Developmental Immunology. 1. 113-125 (1990)

[文献書誌] Takaki,S.: "Molecular cloning and expression of the murine interleukinー5 receptor." The EMBO Journal. 9. 4367-4374 (1990)

[文献書誌] Murata,Y.: "Interleukin 5 and interleukin 3 induce serine and tyrosine phosphorylations of several cellular proteins in an interleukin 5 dependent cell line." Biochemical and Biophysical Recearch Communications. 173. 1102-1108 (1990)

[文献書誌] Yokota,T.: "Handbook of Experimental Pharmacology,Vol.95/I Peptide Growth.Factors and Their Receptors I" SpringerーVerlag, 31 (1990)

[文献書誌] Hirano,T.,and T.Kishimoto: "Handbook of Experimental Pharmacology vol.95/I “Peptide Growth Factors and Their Receptors"" SpringerーVerlag, 35 (1990)

1990 年度実績報告書

新井賢一東京大学, 医科学研究所, 教授 (00012782)