1991 Fiscal Year Annual Research Report

増殖因子による遺伝子発現とDNA複製開始に至るシグナル伝遺機構の研究

Research Project

Project/Area Number	02404086
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	DNA複製 / Gタンパク質 / シグナル伝達 / プライモソ-ム / キナ-ゼ / サイトカイン / 転写因子 / 受容体
Research Abstract	本研究は細胞周期のGIからS期移行の細胞内制御機構について原核細胞の知見をふまえて哺乳動物細胞、特にリンパ球及び血球細胞において検討することを目的とした。1)大腸菌染色体の複製制御:複製開始に関与する、priA依在性φX174型及びdnaA依存性ABC型のプライモソ-ムの機能について以下の点を明らかにした。ColEl型プラスミドの複製において2つのプライモソ-ムは機能的に等価である。また、プライモソ-ム構築部位pasを起点近傍にもつレプリコン(F,R6K,Rtsl)の複製はdnaA要求性で、priA要求性のφX174型プライモソ-ムは必須ではない。一方、dnaAとoriCに依在しない大腸菌染色体のrecA依存性安定DNA合成にはpriA依存性のφX174型プライモソ-ムが関与する。以上から、大腸菌ではABC型あるいはφX174型プライモソ-ムに依存する2種類のレプリコンに大別されることを提唱した。2)酵母細胞のG1/S遷移の制御:DNA複製の開始を負に制御する性フェロモンの細胞内シグナルの伝達の鍵となるG蛋白質のサブユニット(STE4とSTE18)の機能を遺伝学的に検討し、両蛋白質が複合体を形成する可能性を示した。現在両蛋白質の複合体が相互作用するエフェクタ-蛋白質を検索中である。また、染色体複製起点(ARS配列)と相互作用する蛋白質を探索し、保存されたコア配列のTーrichー本鎖DNAに特異的に結合する一連の蛋白質を同定し、その中の100kDa蛋白質をほぼ均一に精製した。現在他の結合蛋白質の精製も進めている。DNA複製の間始を制御すると考えられるCDC7キナ-ゼを増産し、抗体を作製し、その機能についても解析を進めている。3)リンパ球及び血球細胞におけるG1/S遷移の制御:血球細胞のG1/S遷移を正の制御するサイトカインILー3及びGMーCSFの受容体は各サイトカイン特異的なα鎖とシグナル伝達に必要な共通のβ鎖の二本鎖から構成されることを明らかにした。現在これらの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。一方、抗原激刺により活性されたリンパ球はG1/S遷移に伴い種々のサイトカイン遺伝子の転写を透導するが、本研究では特にGMーCSFの発現に関与する転写因子NFーGM2を同定精製し、それがNFーkB/relのfamilyに属することを明らかにした。また、ILー3とILー4についても転写因子を同定単離した。

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ー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ーCSF promoter:nuclear factors that interact with an element shared by three lymphokine genes:GMーCSF,ILー4,and ILー5" Molecular and Cellular Biology. 11. 5894-5909 (1991)
[Publications] Kitamura,T.et.al.: "Exprssion cloning of human ILー3 receptor cDNA and reconstitution of high affinity receptors for human ILー3 and GMーCSF" Cell. 66. 1165-1174 (1991)
[Publications] Tsuboi,A.et.al.: "A nuclear factor NFGM2,essential for induction of GMーCSF gene in response to Tーcell activation:purification from T cell leukemia line Jurkat cell and relationship to NFー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" International Immunology. 3. 245-254 (1991)
[Publications] Watanabe,S.et.al.: "Colony stmulating factors and cytokine receptor network" Curren Opinion in Biotechnology. 2. 227-237 (1991)
[Publications] Kitamura,T.et.al.: "Reconstitution of functional receptors for human granulocyteー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ー3 receptor:temperatureーdependent modulation of AIC2A structure" International Immunology. 3. 1231-1242 (1991)
[Publications] Ozu,M.et.al.: "Regulation of the ILー3 gene and structure and function of the ILー3 receptor" Advance in Neuroimmunology. 2. 32-52 (1992)
[Publications] Wang,H.M.et.al.: "Structure of mouse ILー3 binding protein (AIC2A) Amino acid residues critical for ILー3 binding" Journal of Biological Chemistry. (1992)
[Publications] Garret,K.P.et.al.: "The Cー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ー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ーbase pair DNA sequence motif unique to human ILー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.: "Moleculoar 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

増殖因子による遺伝子発現とDNA複製開始に至るシグナル伝遺機構の研究

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ー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ーCSF promoter:nuclear factors that interact with an element shared by three lymphokine genes:GMーCSF,ILー4,and ILー5" Molecular and Cellular Biology. 11. 5894-5909 (1991)

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

[Publications] Tsuboi,A.et.al.: "A nuclear factor NFGM2,essential for induction of GMーCSF gene in response to Tーcell activation:purification from T cell leukemia line Jurkat cell and relationship to NFー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" International Immunology. 3. 245-254 (1991)

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

[Publications] Kitamura,T.et.al.: "Reconstitution of functional receptors for human granulocyteー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ー3 receptor:temperatureーdependent modulation of AIC2A structure" International Immunology. 3. 1231-1242 (1991)

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

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

[Publications] Garret,K.P.et.al.: "The Cー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ー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ーbase pair DNA sequence motif unique to human ILー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.: "Moleculoar 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)