1991 Fiscal Year Annual Research Report

サイトカインによる細胞の増殖分化の制御とシグナル伝達

Research Project

Project/Area Number	03152034
Research Institution	The University of Tokyo
Principal Investigator	新井賢一東京大学, 医科学研究所, 教授 (00012782)
Co-Investigator(Kenkyū-buntansha)	村松正明東京大学, 医科学研究所, 教務職員 (50230008) 正井久雄東京大学, 医科学研究所, 助手 (40229349) 中山直樹東京大学, 医科学研究所, 助手 (80227967) 横田崇東京大学, 医科学研究所, 助教授 (50134622)
Keywords	サイトカイン / 遺伝子発現 / 転写因子 / シグナル伝達 / 受容体 / T細胞
Research Abstract	本年度はサイトカインによるリンパ球及び血球細胞のG1/S遷移機構について、T細胞の活性化に伴うサイトカイン遺伝子の発現誘導のメカニズムとサイトカイン受容体の解析を通して検討することを目的とした。1)サイトカイン遺伝子発現誘導に必要な転写因子の単離:活性化されたT細胞はG1/S遷移し、その際種々のサイトカイン遺伝子の転写が誘発される。本研究ではGMーCSFの遺伝子発現誘導に関与する領域CLE2/GCboxに活性化特異的に結合する転写因子NFーGM2を活性化T細胞の抽出液中に同定し、精製し、同因子がNFーkB/relのfamilyに属することを明らかにした。そのcDNAの候補もSouthーWestern法によりクロ-ニングした。さらに、ILー3のGCboxに結合する転写因子を同定し、やはりSouthーWestern法によりその候補を直接cDNAとして単離した。現在これらのクロ-ンの解析を進めている。2)ILー3/GMーCSF受容体はβ鎖を共有する:cDNAクロ-ニングの結果、血液幹細胞のG1/S遷移と分化へのcommitmentを制御するサイトカイン,ILー3及びGMーCSFの受容体は各々に特異的で低親和性で結合する能力を有するα鎖と高親和性の発現には必要であるが自身では必ずしもサイトカインと結合しないβ鎖の二本鎖から構成されることを明らかにした。両鎖cDNAの繊維芽細胞及びT細胞へのトランスフェクションにより、各サイトカイン受容体は一種類のβ鎖を共有すること(マウスのAIC2b、及びヒトのKH97)、しかもこのβ鎖がサイトカインのシグナル伝達に必須であることを見いだした。各サイトカインの効果は標的細胞により異なるため、作用の違いをシグナル転換機構のレベルで説明するため、現在第3の鎖の存在の検討及び各受容体が相互作用するチロシンキナ-ゼの解析を進めている。又、受容体各鎖の遺伝子の構造と染色体上の位置についても解析を行なっている。

Research Products
(19 results)

All Other

All Publications (19 results)

[Publications] Mauxion,F.et.al.: "Comparison of constitutive and inducible transcriptional enhancement mediated by KB-related sequences:modulation of activity in B cells by human T cell leukemia virus type I tax gene" Proceedings of National Academy of Science USA. 88. 2141-2145 (1991)
[Publications] Miyatake,S.et.al.: "Characterization of the mouse GM-CSF promoter:nuclear factors that interact with an element shared by three lymphokine genes:GM-CSF,IL-4,and IL-5" Molecular and Cellular Biology. 11. 5894-5909 (1991)
[Publications] Kitamura,T.et.al.: "Expression cloning of human IL-3 receptor cDNA and reconstitution of high affinity receptors for human IL-3 and GM-CSF" Cell. 66. 1165-1174 (1991)
[Publications] Tsuboi,A.et.al.: "A nuclear factor NFGM2,essential for induction of GM-CSF gene in response to T-cell activation:purification from T cell leukemia line Jurkat cell and relationship to NF-kB" International Immunology. 3. 807-817 (1991)
[Publications] Nomura,N.et.al.: "Identification of eleven single strand initiation sequences (ssi) for priming of DNA replication in the F,R6K,R100 and ColE2 plasmids" Gene. 108. 15-22 (1991)
[Publications] Arai,K.et.al.: "Cytokine network and regulation of inflammatory response" Excepta Medica. (1991)
[Publications] Nishida,J.et.al.: "Definition of GC rich motif as regulatoy sequence of thehuman IL-3 gene:coordinate regulation of the IL-3 gene by CLE2/GC box of the GM-CSF gene in T cell activation" International Immunology. 3. 245-254 (1991)
[Publications] Watanabe,S.et.al.: "Colony stimulating factors and cytokine receptor network" Current Opinion in Biotechnology. 2. 227-237 (1991)
[Publications] Kitamura,T.et.al.: "Reconstitution of functional receptors for human granulocyte-macrophage colony stimulating factor:requirement of the b subunit for signaling" Proceedings of National Academy of Science USA. 88. 5082-5086 (1991)
[Publications] Schreurs,J.et.al.: "AIC2A is a component of the purified high affinity mouse IL-3 receptor:temperature-dependent modulation of AIC2A structure" International Immunology. 3. 1231-1242 (1991)
[Publications] Ozu,M.et.al.: "Regulation of the IL-3 gene and structure and function of the IL-3 receptor" Advance in Neuroimmunology. 2. 32-52 (1992)
[Publications] Wang,H.M.et.al.: "Structure of mouse IL-3 binding protein (AIC2A):Amino acid residues critical for IL-3 binding" Journal of Biological Chemistry. (1992)
[Publications] Garret,K.P.et.al.: "The C-terminus of rat RAP30 is similar in sequence to region 4 of bacterial sigma factors and is required for function" submitted. (1992)
[Publications] Hisatsune,T.et.al.: "A suppressive lymphokine derived from Ts clone 13G2 is IL-10" Lymphokine Research. (1992)
[Publications] Miyajima A.et.al.: "Cytokine receptors and signal transduction" Annual Review of Immunology. 10. 295-331 (1992)
[Publications] Abe,E.et.al.: "Eleven-base pair DNA sequence motif unique to human IL-4 gene confers responsiveness to T cell activation signals" Proceedings of National Academy of Science USA. (1992)
[Publications] Hatake,K.et.al.: "Human Cytokines" Blackwell Scientific Publications Inc.(Aggarwal,B.B.and Gutterman,J.U.,eds), 135-142(8) (1991)
[Publications] Miyatake,S.et.al.: "Molecular Biology of Lymphokines" VCH Publishers,Inc., 35-37(3) (1991)
[Publications] Yokota,T.et.al.: "Cytokines and Infectious Disease" Marcel Decker Inc.(Bermudes,L.Z.and Steems,P.,eds), (1992)

1991 Fiscal Year Annual Research Report

サイトカインによる細胞の増殖分化の制御とシグナル伝達

Principal Investigator

新井 賢一 東京大学, 医科学研究所, 教授 (00012782)

Research Products

[Publications] Mauxion,F.et.al.: "Comparison of constitutive and inducible transcriptional enhancement mediated by KB-related sequences:modulation of activity in B cells by human T cell leukemia virus type I tax gene" Proceedings of National Academy of Science USA. 88. 2141-2145 (1991)

[Publications] Miyatake,S.et.al.: "Characterization of the mouse GM-CSF promoter:nuclear factors that interact with an element shared by three lymphokine genes:GM-CSF,IL-4,and IL-5" Molecular and Cellular Biology. 11. 5894-5909 (1991)

[Publications] Kitamura,T.et.al.: "Expression cloning of human IL-3 receptor cDNA and reconstitution of high affinity receptors for human IL-3 and GM-CSF" Cell. 66. 1165-1174 (1991)

[Publications] Tsuboi,A.et.al.: "A nuclear factor NFGM2,essential for induction of GM-CSF gene in response to T-cell activation:purification from T cell leukemia line Jurkat cell and relationship to NF-kB" International Immunology. 3. 807-817 (1991)

[Publications] Nomura,N.et.al.: "Identification of eleven single strand initiation sequences (ssi) for priming of DNA replication in the F,R6K,R100 and ColE2 plasmids" Gene. 108. 15-22 (1991)

[Publications] Arai,K.et.al.: "Cytokine network and regulation of inflammatory response" Excepta Medica. (1991)

[Publications] Nishida,J.et.al.: "Definition of GC rich motif as regulatoy sequence of thehuman IL-3 gene:coordinate regulation of the IL-3 gene by CLE2/GC box of the GM-CSF gene in T cell activation" International Immunology. 3. 245-254 (1991)

[Publications] Watanabe,S.et.al.: "Colony stimulating factors and cytokine receptor network" Current Opinion in Biotechnology. 2. 227-237 (1991)

[Publications] Kitamura,T.et.al.: "Reconstitution of functional receptors for human granulocyte-macrophage colony stimulating factor:requirement of the b subunit for signaling" Proceedings of National Academy of Science USA. 88. 5082-5086 (1991)

[Publications] Schreurs,J.et.al.: "AIC2A is a component of the purified high affinity mouse IL-3 receptor:temperature-dependent modulation of AIC2A structure" International Immunology. 3. 1231-1242 (1991)

[Publications] Ozu,M.et.al.: "Regulation of the IL-3 gene and structure and function of the IL-3 receptor" Advance in Neuroimmunology. 2. 32-52 (1992)

[Publications] Wang,H.M.et.al.: "Structure of mouse IL-3 binding protein (AIC2A):Amino acid residues critical for IL-3 binding" Journal of Biological Chemistry. (1992)

[Publications] Garret,K.P.et.al.: "The C-terminus of rat RAP30 is similar in sequence to region 4 of bacterial sigma factors and is required for function" submitted. (1992)

[Publications] Hisatsune,T.et.al.: "A suppressive lymphokine derived from Ts clone 13G2 is IL-10" Lymphokine Research. (1992)

[Publications] Miyajima A.et.al.: "Cytokine receptors and signal transduction" Annual Review of Immunology. 10. 295-331 (1992)

[Publications] Abe,E.et.al.: "Eleven-base pair DNA sequence motif unique to human IL-4 gene confers responsiveness to T cell activation signals" Proceedings of National Academy of Science USA. (1992)

[Publications] Hatake,K.et.al.: "Human Cytokines" Blackwell Scientific Publications Inc.(Aggarwal,B.B.and Gutterman,J.U.,eds), 135-142(8) (1991)

[Publications] Miyatake,S.et.al.: "Molecular Biology of Lymphokines" VCH Publishers,Inc., 35-37(3) (1991)

[Publications] Yokota,T.et.al.: "Cytokines and Infectious Disease" Marcel Decker Inc.(Bermudes,L.Z.and Steems,P.,eds), (1992)

新井賢一東京大学, 医科学研究所, 教授 (00012782)