1998 Fiscal Year Annual Research Report

TGF-β-DPC-4による細胞周期制御機構の解析

Research Project

Project/Area Number	09470042
Research Institution	The University of Tokyo
Principal Investigator	秋山徹東京大学, 分子細胞生物学研究所, 教授 (70150745)
Keywords	DPC4 / TGF-β / 標的遺伝子 / one-hybrid / サブトラクション / 細胞周期
Research Abstract	DPC4(SMAD4)はSMAD2あるいはSMAD3と複合体を形成し、TGF-P標的遺伝子のプロモーター領域に直接結合することで、標的遺伝子の発現を制御する。そこで、DNAと直接相互作用するDPC4をbaitに用いたyeast one-hybrid法を行うことでDPC4の標的遺伝子の探索を行った(one-hybrid法はbaitタンパク質と特異的に相互作用するDNA断片を単離する方法)。約2x10^5種類のヒトゲノム由来DNA断片をスクリーニングした結果、3種類のDNA断片を得た。One-hybrid法で得られたDNA断片内にはDPC4が認識するコンセンサス配列が実際に含まれており、本方法が標的遺伝子の探索に有効な方法のひとつであることが確認された。塩基配列決定後のデータベース検索により、1つがヒト28S rRNA遺伝子と一致することがわかった。残る2つのDNA断片については、目下解析中である。DPC4標的遺伝子の探索の別のアプローチとして、サブトラクション法及びプロモーター・トラップ法によるスクリーニングも進めており、現在、細胞周期・シグナル伝達・細胞骨格関連の遺伝子群と新規遺伝子群に焦点を定めた解析を行っている。さらに、DPC4による細胞周期制御機構を明らかにする目的でDPC4に特異的に結合する新規蛋白質の探索も昨年度に引き続き行っている。

Research Products

(13 results)

All Other

All Publications (13 results)

[Publications] T.Nakamura et al.: "Axin,an inhibitor of the Wnt signaling pathway,interacts with β-catenin,CSK-3β and APC and reduces the β-catenin level." Genes Cells. 3. 395-403 (1998)
[Publications] T.Senda et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)
[Publications] K.Inoue et al.: "Wilms'tumor gene(WT1)competes with differentiation-inducing signal in hematopoietic progenitor cells." Blood. 91. 2969-2976 (1998)
[Publications] H.Nakagawa et al.: "Identification of a brain-specific APC homologue,APCL,and its interaction with β-catenin." Cancer Res.58. 5176-5181 (1998)
[Publications] Y.Dabashi et al.: "CDK4,a possible critical regulator of apoptosis in rat pheochromocytoma PC12 cells." Biochem.Biophys.Res.Commun.253. 609-613 (1998)
[Publications] T.Yamagami et al.: "Suppression of Wilms'tumor gene(WT1)expression induces G2/M arrest in leukemic cells." Leuk.Res.22. 383-384 (1998)
[Publications] K.Mori et al.: "Identification of brain-specific splicing variants of the hDLG1 gene and altered splicing in neuroblastoma cell lines." J.Hum.Genet.43. 123-127 (1998)
[Publications] T.Yamochi et al.: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL." Oncogene. 18. 487-494 (1999)
[Publications] T.Tezuka et al.: "PSD-95 promotes Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunit NR2A." Proc.Natl.Acad.Sci.USA96. 435-440 (1999)
[Publications] F.Hamada et al.: "Negative regulation of Wingless signaling by D-Axin,a Drosophila homologue of Axin." Science. 283. 1739-1742
[Publications] T.Senda et al.: "Association of the MCC(Mutated in colorectal cancer)gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy. (in press).
[Publications] M.Takemura et al.: "Nucleolar protein B23.1 binds to retinoblastoma protein and synergistically stimulates DNA polymerase alpha activity." J.Biochem.(in press).
[Publications] R.B.Chandra et al.: "Localization of the GAP family SPAL in the Rat Esophagus and Heart." Medical Electronmicroscopy. (in press).

1998 Fiscal Year Annual Research Report

TGF-β-DPC-4による細胞周期制御機構の解析

Principal Investigator

秋山 徹 東京大学, 分子細胞生物学研究所, 教授 (70150745)

Research Products

[Publications] T.Nakamura et al.: "Axin,an inhibitor of the Wnt signaling pathway,interacts with β-catenin,CSK-3β and APC and reduces the β-catenin level." Genes Cells. 3. 395-403 (1998)

[Publications] T.Senda et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)

[Publications] K.Inoue et al.: "Wilms'tumor gene(WT1)competes with differentiation-inducing signal in hematopoietic progenitor cells." Blood. 91. 2969-2976 (1998)

[Publications] H.Nakagawa et al.: "Identification of a brain-specific APC homologue,APCL,and its interaction with β-catenin." Cancer Res.58. 5176-5181 (1998)

[Publications] Y.Dabashi et al.: "CDK4,a possible critical regulator of apoptosis in rat pheochromocytoma PC12 cells." Biochem.Biophys.Res.Commun.253. 609-613 (1998)

[Publications] T.Yamagami et al.: "Suppression of Wilms'tumor gene(WT1)expression induces G2/M arrest in leukemic cells." Leuk.Res.22. 383-384 (1998)

[Publications] K.Mori et al.: "Identification of brain-specific splicing variants of the hDLG1 gene and altered splicing in neuroblastoma cell lines." J.Hum.Genet.43. 123-127 (1998)

[Publications] T.Yamochi et al.: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL." Oncogene. 18. 487-494 (1999)

[Publications] T.Tezuka et al.: "PSD-95 promotes Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunit NR2A." Proc.Natl.Acad.Sci.USA96. 435-440 (1999)

[Publications] F.Hamada et al.: "Negative regulation of Wingless signaling by D-Axin,a Drosophila homologue of Axin." Science. 283. 1739-1742

[Publications] T.Senda et al.: "Association of the MCC(Mutated in colorectal cancer)gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy. (in press).

[Publications] M.Takemura et al.: "Nucleolar protein B23.1 binds to retinoblastoma protein and synergistically stimulates DNA polymerase alpha activity." J.Biochem.(in press).

[Publications] R.B.Chandra et al.: "Localization of the GAP family SPAL in the Rat Esophagus and Heart." Medical Electronmicroscopy. (in press).

秋山徹東京大学, 分子細胞生物学研究所, 教授 (70150745)