1999 Fiscal Year Annual Research Report

転写因子間の相互作用と機能発現の分子機構

Research Project

Project/Area Number	09277102
Research Institution	Tohoku University
Principal Investigator	藤井義明東北大学, 大学院・理学研究科, 教授 (00098146)
Co-Investigator(Kenkyū-buntansha)	大熊芳明大阪大学, 細胞生体工学センター, 助教授 (70192515) 上田均国立遺伝学研究所, 形質遺伝研究室, 助手 (60201349) 審良静男大阪大学, 微生物病研究所, 教授 (50192919) 半田宏東京工業大学, フロンティア創造共同研究センター, 教授 (80107432) 萩原正敏東京医科歯科大学, 難治疾患研究所, 教授 (10208423)
Keywords	転写共役因子 / 転写因子 / 転写開始複合体 / リン酸化 / 転写伸長反応 / スプライシング / DNA結合能 / 相互作用
Research Abstract	本計画研究班の研究課題は転写共役因子も含めて、転写因子間の相互作用が最終的に遺伝子発現としてどう結果するかというメカニズムを分子のレベルで解明することを主な目的としている。久武は、高次複合体を形成して働くTFIIH、TFIID、PolIIを組み換えDNAの方法で生合成した構成成分で再構築して、転写における各成分の役割を研究している。現在、TFIIHの9サブユニットからなる再構成に成功して、ERCC3のヘリカーゼ活性が転写には必須であることを明らかにした。大熊は、転写開始複合体内でTFIIFのみがPolIIをリン酸化することができ、2位と5位のSerがリン酸化される。このリン酸化は転写に重要な役割を果たしているがSer-5のリン酸化にTFIIEが必要であることを示した。また、TFIIEサブユニットTFIIEbのコアドメインの立体構造を西村と決定した。半田は、転写伸長反応を正と負にコントロールする因子がp-TEFbとDSIFであることを明らかにし、DSIFの構造サブユニットの構造をcDNAクローニングによって解明した。さらに、転写に阻害的に働くNELFを発見し、精製して5つのサブユニットから成ることを示した。萩原は、RNAのスプライシングに働くSF2やS35などのRSドメインのリン酸化酵素として、SRPK1の他にSRPK2を見つけ、SF2/ASFがリン酸化に伴って核外に移行することを見つけた。堀越は、p300のHATドメインがSp1のDNA結合ドメインと相互作用してDNA結合能を促進することを見出した。この他に、TFIIDと相互作用する因子を数多く単離しており、それらの機能の解析をしている。原と上田は共同して、FTZ-F1とその共役因子FTZとがLXXLLモチーフで結合し、ftz遺伝子の発現に関与していることを示した。この他に、原は、PPARのニワトリのホモログを3種分離して、リガンドとの結合特異性が一つのアミノ酸の置換によって決められることを示した。上田は、FTZ-F1の共役因子MBF1に相互作用する因子APTを分離して、そのbZip領域がその相互作用部位であることを決めた。加藤は、エストロゲン受容体のAF1領域の共役因子として68KDのRNAヘリカーゼ結合タンパクとDEADboxを持つRNA結合タンパク質p72を同定した。審良は、Cre-loxp系を用いてヒフ及び乳腺特異的にSTAT3を欠損するマウスを作製し、STAT欠損マウスではヒフの分化には異常は認められないが、創傷治癒の遅延を認めた。その原因は、STAT3欠損ケラチノサイトが正常細胞に比べて、細胞移動が障害されているためであることが判った。藤井は、AhR欠失マウスを用いてAhRがベンツピレンによるヒフ癌はAhRが関与していることを示した。さらに、低酸素に応じて遺伝子を活性化する転写因子HIF-1αとHLFは、低酸素によるタンパク質の安定化による増量とC末端の転写活性化ドメインにあるCysの酸化還元シグナルによる還元によって活性化されることを明らかにした。

Research Products
(25 results)

All Other

All Publications (25 results)

[Publications] J.Mimura, et.al: "Identification of a novel mechanism of regulation of Ah (dioxin) receptor function"Genes Dev.. 13. 20-25 (1999)
[Publications] M.Ema, et.al: "Molecular mechanism of transcription activation by HLF and HIF1 a in response to hypoxia : their stabilization and redox signal induced interaction with CBP/p300"EMBO J.. 18. 1905-1914 (1999)
[Publications] R.J.Denver, et.al: "Basic Transcription Element Binding Protein (BTEB) is a Thyroid Hormone-Regulated Gene in the Developing Ventral Nervous System : Evidence for a Role in Neurite Outgrowth"J. Biol. Chem.. 274. 23128-23134 (1999)
[Publications] S.Sano, et.al: "Kerationocyte-specific ablation of Stat3 exhibits impaired skin remodeling but does not affect skin morphogenesis"EMBO J.. 18. 4657-4668 (1999)
[Publications] T.Kaisho, et.al: "Impaired natural killer cytotoxic activity and interferon-g production in C/EBPg-deficient mice"J. Exp. Med.. 190. 1573-1581 (1999)
[Publications] K.Hoshino, et.al: "Absence of IL-1 receptor-related T1/ST2 does not affect Th2 cell development and its effector function"J. Exp. Med.. 190. 1541-1547 (1999)
[Publications] Y. Fujii, et.al: "Crystal structure of an IRF/DNA complex reveals a novel DNA recognition and cooperactive binding to a tandem repeat of the core sequence"EMBO J.. 18. 5028-5041 (1999)
[Publications] R.Maesaki, et.al: "The structural basis of Rho effector recognition revealed by the Crystal structure of human RhoA complexed with the effector domain of PKN/PRK1"Molecular Cell. 4. 793-803 (1999)
[Publications] S.Hirotsu, et.al: "Crystal structure of a stress-induced multifunctional 2-Cys peroxiredoxin, heme-binding protein 23kDa/proliferation-associated gene product"Proc. Natl. Acad. Sci. USA. 96. 12333-12338 (1999)
[Publications] Y. Nagai, et.al: "Detction of cAMP-induced phosphorylation with a novel fluorescent indicator"Nat.Biotechnol.. (in press). (2000)
[Publications] H.Sakaguchi, et.al: "Song-Induced Phosphorylation of cAMP Response Element Binding Protein in the Songbird Brain"J.Neuroscience. 19. 3973-3981 (1999)
[Publications] J.Koizumi, et.al: "The subcellular localization of SF2/ASF is regulated by the direct interaction with SR protein kinases(SRPKs)"J. Biol. Chem.. 274. 11125-11131 (1999)
[Publications] M.Kobayashi, et.al: "Identification of a photoreceptor cell-specific nuclear receptor"Proc. Natl. Acad. Sci. USA. 96. 4814-4819 (1999)
[Publications] Y.Yamaguchi, et.al: "NELF, a multiple complex containing RD, cooperates with DSIF to repress RNA polymerase II elongation"Cell. 97. 41-51 (1999)
[Publications] Y.Yamaguchi, et.al: "Structure and function of the human transcription elongation factor DSIF"J. Biol. Chem.. 274. 8085-8092 (1999)
[Publications] J-I.Sawada, et.al: "Synergistic trascriptional activation of hGABP and select members of the ATF/CREB family"J. Biol. Chem.. 274. 35475-35482 (1999)
[Publications] K.Imai, et.al: "The fission yeast rpal7+ gene encodes a functional homolog of AC19, a subunit of RNA polymerase I and III of Saccharomyces cerevisiae"Mol. Gen. Genet.. 261. 364-373 (1999)
[Publications] Y.Imazawa, et.al: "Isolation and characterization of the fission yeast gene rpa42+ which encodes a subunit shared by RNA polymerase I and III"Mol. Gen. Genet.. 262. 794-757 (1999)
[Publications] A.Ishiguro, et.al: "The Rpb6 Subunit of Fission Yeast RNA Polymerase II Is a Contact Target of the Transcription Elongation Factor TFIIS"Mol. Cell. Biol.. 20. 1263-1270 (2000)
[Publications] R.Maesaki, et.al: "The structural basis of Rho effector recognition revealed by the Crystal structure of human RhoA complexed with the effector domain of PKN/PRK1"Molecular Cell. 4. 793-803 (1999)
[Publications] K.Sekine, et.al: "FGF10 is essential for the limb and lung formation. Nature Genetics"Nature Genetics. 21. 138-141 (1999)
[Publications] K.Takeyama, et.al: "Selective interaction of vitamin D receptor with transcriptional coactivators by a vitamin D analog"Mol. Cell. Biol.. 19. 1049-1055 (1999)
[Publications] J. Yanagisawa, et.al: "Convergence of TGFb and vitamin D signaling pathways on SMAD proteins acting as common transcriptional co-activators"Science. 283. 1317-1321 (1999)
[Publications] R.Himukai, et.al: "Relationship between subcellular localization and structure of catalytic domains of FKBP-type PPIases"J. Biochem. 126. 879-888 (1999)
[Publications] T.Suzuki, et.al: "Regulation of interaction of the acetyltransferase region of p300 and the DNA-binding domain of Sp1 on and through DNA binding"Genes Cells. 5. 29-42 (2000)

1999 Fiscal Year Annual Research Report

転写因子間の相互作用と機能発現の分子機構

Principal Investigator

藤井 義明 東北大学, 大学院・理学研究科, 教授 (00098146)

Research Products

[Publications] J.Mimura, et.al: "Identification of a novel mechanism of regulation of Ah (dioxin) receptor function"Genes Dev.. 13. 20-25 (1999)

[Publications] M.Ema, et.al: "Molecular mechanism of transcription activation by HLF and HIF1 a in response to hypoxia : their stabilization and redox signal induced interaction with CBP/p300"EMBO J.. 18. 1905-1914 (1999)

[Publications] R.J.Denver, et.al: "Basic Transcription Element Binding Protein (BTEB) is a Thyroid Hormone-Regulated Gene in the Developing Ventral Nervous System : Evidence for a Role in Neurite Outgrowth"J. Biol. Chem.. 274. 23128-23134 (1999)

[Publications] S.Sano, et.al: "Kerationocyte-specific ablation of Stat3 exhibits impaired skin remodeling but does not affect skin morphogenesis"EMBO J.. 18. 4657-4668 (1999)

[Publications] T.Kaisho, et.al: "Impaired natural killer cytotoxic activity and interferon-g production in C/EBPg-deficient mice"J. Exp. Med.. 190. 1573-1581 (1999)

[Publications] K.Hoshino, et.al: "Absence of IL-1 receptor-related T1/ST2 does not affect Th2 cell development and its effector function"J. Exp. Med.. 190. 1541-1547 (1999)

[Publications] Y. Fujii, et.al: "Crystal structure of an IRF/DNA complex reveals a novel DNA recognition and cooperactive binding to a tandem repeat of the core sequence"EMBO J.. 18. 5028-5041 (1999)

[Publications] R.Maesaki, et.al: "The structural basis of Rho effector recognition revealed by the Crystal structure of human RhoA complexed with the effector domain of PKN/PRK1"Molecular Cell. 4. 793-803 (1999)

[Publications] S.Hirotsu, et.al: "Crystal structure of a stress-induced multifunctional 2-Cys peroxiredoxin, heme-binding protein 23kDa/proliferation-associated gene product"Proc. Natl. Acad. Sci. USA. 96. 12333-12338 (1999)

[Publications] Y. Nagai, et.al: "Detction of cAMP-induced phosphorylation with a novel fluorescent indicator"Nat.Biotechnol.. (in press). (2000)

[Publications] H.Sakaguchi, et.al: "Song-Induced Phosphorylation of cAMP Response Element Binding Protein in the Songbird Brain"J.Neuroscience. 19. 3973-3981 (1999)

[Publications] J.Koizumi, et.al: "The subcellular localization of SF2/ASF is regulated by the direct interaction with SR protein kinases(SRPKs)"J. Biol. Chem.. 274. 11125-11131 (1999)

[Publications] M.Kobayashi, et.al: "Identification of a photoreceptor cell-specific nuclear receptor"Proc. Natl. Acad. Sci. USA. 96. 4814-4819 (1999)

[Publications] Y.Yamaguchi, et.al: "NELF, a multiple complex containing RD, cooperates with DSIF to repress RNA polymerase II elongation"Cell. 97. 41-51 (1999)

[Publications] Y.Yamaguchi, et.al: "Structure and function of the human transcription elongation factor DSIF"J. Biol. Chem.. 274. 8085-8092 (1999)

[Publications] J-I.Sawada, et.al: "Synergistic trascriptional activation of hGABP and select members of the ATF/CREB family"J. Biol. Chem.. 274. 35475-35482 (1999)

[Publications] K.Imai, et.al: "The fission yeast rpal7+ gene encodes a functional homolog of AC19, a subunit of RNA polymerase I and III of Saccharomyces cerevisiae"Mol. Gen. Genet.. 261. 364-373 (1999)

[Publications] Y.Imazawa, et.al: "Isolation and characterization of the fission yeast gene rpa42+ which encodes a subunit shared by RNA polymerase I and III"Mol. Gen. Genet.. 262. 794-757 (1999)

[Publications] A.Ishiguro, et.al: "The Rpb6 Subunit of Fission Yeast RNA Polymerase II Is a Contact Target of the Transcription Elongation Factor TFIIS"Mol. Cell. Biol.. 20. 1263-1270 (2000)

[Publications] R.Maesaki, et.al: "The structural basis of Rho effector recognition revealed by the Crystal structure of human RhoA complexed with the effector domain of PKN/PRK1"Molecular Cell. 4. 793-803 (1999)

[Publications] K.Sekine, et.al: "FGF10 is essential for the limb and lung formation. Nature Genetics"Nature Genetics. 21. 138-141 (1999)

[Publications] K.Takeyama, et.al: "Selective interaction of vitamin D receptor with transcriptional coactivators by a vitamin D analog"Mol. Cell. Biol.. 19. 1049-1055 (1999)

[Publications] J. Yanagisawa, et.al: "Convergence of TGFb and vitamin D signaling pathways on SMAD proteins acting as common transcriptional co-activators"Science. 283. 1317-1321 (1999)

[Publications] R.Himukai, et.al: "Relationship between subcellular localization and structure of catalytic domains of FKBP-type PPIases"J. Biochem. 126. 879-888 (1999)

[Publications] T.Suzuki, et.al: "Regulation of interaction of the acetyltransferase region of p300 and the DNA-binding domain of Sp1 on and through DNA binding"Genes Cells. 5. 29-42 (2000)

藤井義明東北大学, 大学院・理学研究科, 教授 (00098146)