2002 Fiscal Year Annual Research Report

膵臓腺房細胞におけるユビキチン様蛋白質の役割

Research Project

Project/Area Number	13670583
Research Institution	Institute for Developmental Research, Aichi Human Service Center
Principal Investigator	稲熊裕愛知県心身障害者コロニー発達障害研究所, 生化学部, 主任研究員 (10250250)
Co-Investigator(Kenkyū-buntansha)	伊東秀記愛知県心身障害者コロニー発達障害研究所, 生化学部, 研究員 (40311443)
Keywords	SUMO / ユビキチン / DNA修復 / APEX / TTRAP / 転写
Research Abstract	我々はこれまでに抗SUMO-1特異精製抗体によるウェスタンブロット解析で、遊離型SUMO-1が膵臓に極めて高い濃度で存在し、マウス胎仔およびヒト膵臓組織の免疫組織化学的検索においても腺房細胞が非常に強い陽性を示すことを確認した。培養細胞MIA PaCa-2でも遊離型SUMO-1が多く存在した。さらに、抗SUMO-2/3特異精製抗体を用いたウェスタンブロット解析でも、遊離型SUMO-2がSUMO-1と同様に膵臓に多く存在した。SUMO-2化された蛋白質は、細胞へのストレス(熱)負荷時、ただちに高分子量化し多くが不溶性分画に移行した。SUMO-2をbaitとしてTwo-hybridスクリーニングを行い、TTRAP (TNF receptor and TRAF associated protein)が相互作用することを見つけた。さらに、抗SUMO-2抗体を作製し、細胞内でSUMO-2とTTRAPが核に共局在することを確認した。TTRAPはN末端側に核移行シグナルが予測され、ここに変異を入れると核への局在が失われた。TTRAPのC末端側にはDNA修復機能やp53、NF-κB、AP-1等の転写因子を還元する機能をもつAPE-1のモチーフが保存されていた。精製したTTRAPはAP-siteに作用したが、8-oxo-dGへの作用は弱かった。AP-siteへの作用はEDTAの添加によって阻止されたことからMg^<2+>依存的な反応であることが示唆された。さらに、TTRAPはUBC9やp53とも相互作用することが明かとなった。我々が見い出した知見は、SUMO修飾が関わる遺伝子修復機構および遺伝子発現調節の解明に新たな展開を与え、膵炎および膵癌発生の予防と機構の解明、膵臓細胞のストレス応答機構の解明に役立つと考えている。

Research Products

(12 results)

All Other

All Publications (12 results)

[Publications] Katoh-Semba, et al.: "Induction of brain-derived neurotrophic factor by convulsant drugs in the rat brain : involvement of region-specific voltage-dependent calcium channels"J Neurochem. 77. 71-83 (2001)
[Publications] Kato, et al.: "Protein kinase inhibitors can suppress stress-induced dissociation of Hsp27"Cell Stress Chaperones. 6. 16-20 (2001)
[Publications] Kato, et al.: "Ser-59 is the major phosphorylation site in alphaB-crystallin accumulated in the brains of patients with Alexander's disease"J Neurochem. 76. 730-736 (2001)
[Publications] Ito, et al.: "Regulation of the levels of small heat-shock proteins during differentiation of C2C12 cells"Exp Cell Res. 266. 213-221 (2001)
[Publications] Ito, et al.: "Phosphorylation-induced change of the oligomerization state of alpha B-crystalline"J Biol Chem. 276. 5346-5342 (2001)
[Publications] Inaguma, et al.: "AlphaB-crystallin phosphorylated at Ser-59 is localized in centrosomes and midbodies during mitosis"Eur J Cell Biol. 80. 741-748 (2001)
[Publications] Ikeda, et al.: "Virological properties and nucleotide sequences of Cas-E-type endogenous ecotropic murine leukemia viruses in South Asian wild mice, Mus musculus castaneus"J Virol. 75. 5049-5058 (2001)
[Publications] Kato, et al.: "Innervation-dependent phosphorylation and accumulation of alphaB-crystallin and Hsp27 as insoluble complexes in disused muscle"FASEB J. 16. 1432-1434 (2002)
[Publications] Katoh-Semba, et al.: "Riluzole enhances expression of brain-derived neurotrophic factor with consequent proliferation of granule precursor cells in the rat hippocampus"FASEB J. 16. 1328-1330 (2002)
[Publications] Ito, et al.: "Inhibition of proteasomes induces accumulation, phosphorylation, and recruitment of HSP27 and alphaB-Crystallin to aggresomes"J Biochem. 131. 593-603 (2002)
[Publications] Kato, et al.: "Progress in Molecular and Subcellular Biology"Expression and phosphorylation of mammalian small heat shock proteins. 22 (2002)
[Publications] 伊東秀記, 他: "日本薬理学雑誌"低分子量HSPと細胞内タンパク質凝集体. 6 (2003)

2002 Fiscal Year Annual Research Report

膵臓腺房細胞におけるユビキチン様蛋白質の役割

Principal Investigator

稲熊 裕 愛知県心身障害者コロニー発達障害研究所, 生化学部, 主任研究員 (10250250)

Research Products

[Publications] Katoh-Semba, et al.: "Induction of brain-derived neurotrophic factor by convulsant drugs in the rat brain : involvement of region-specific voltage-dependent calcium channels"J Neurochem. 77. 71-83 (2001)

[Publications] Kato, et al.: "Protein kinase inhibitors can suppress stress-induced dissociation of Hsp27"Cell Stress Chaperones. 6. 16-20 (2001)

[Publications] Kato, et al.: "Ser-59 is the major phosphorylation site in alphaB-crystallin accumulated in the brains of patients with Alexander's disease"J Neurochem. 76. 730-736 (2001)

[Publications] Ito, et al.: "Regulation of the levels of small heat-shock proteins during differentiation of C2C12 cells"Exp Cell Res. 266. 213-221 (2001)

[Publications] Ito, et al.: "Phosphorylation-induced change of the oligomerization state of alpha B-crystalline"J Biol Chem. 276. 5346-5342 (2001)

[Publications] Inaguma, et al.: "AlphaB-crystallin phosphorylated at Ser-59 is localized in centrosomes and midbodies during mitosis"Eur J Cell Biol. 80. 741-748 (2001)

[Publications] Ikeda, et al.: "Virological properties and nucleotide sequences of Cas-E-type endogenous ecotropic murine leukemia viruses in South Asian wild mice, Mus musculus castaneus"J Virol. 75. 5049-5058 (2001)

[Publications] Kato, et al.: "Innervation-dependent phosphorylation and accumulation of alphaB-crystallin and Hsp27 as insoluble complexes in disused muscle"FASEB J. 16. 1432-1434 (2002)

[Publications] Katoh-Semba, et al.: "Riluzole enhances expression of brain-derived neurotrophic factor with consequent proliferation of granule precursor cells in the rat hippocampus"FASEB J. 16. 1328-1330 (2002)

[Publications] Ito, et al.: "Inhibition of proteasomes induces accumulation, phosphorylation, and recruitment of HSP27 and alphaB-Crystallin to aggresomes"J Biochem. 131. 593-603 (2002)

[Publications] Kato, et al.: "Progress in Molecular and Subcellular Biology"Expression and phosphorylation of mammalian small heat shock proteins. 22 (2002)

[Publications] 伊東秀記, 他: "日本薬理学雑誌"低分子量HSPと細胞内タンパク質凝集体. 6 (2003)

稲熊裕愛知県心身障害者コロニー発達障害研究所, 生化学部, 主任研究員 (10250250)