2001 年度実績報告書

外来異物に対する信号伝達機構を利用した環境モニター動物の開発

研究課題

研究課題/領域番号	12557015
研究機関	筑波大学
研究代表者	山本雅之筑波大学, 基礎医学系, 教授 (50166823)
研究分担者	小林麻己人筑波大学, 基礎医学系, 講師 (50254941)
キーワード	親電子性物質 / 異物代謝 / 転写制御 / バイオモニター / Nrf2 / 酸化ストレス / マウス / ゼブラフィッシュ
研究概要	毒性化学物質のバイオモニター系開発を目的に,外来異物に対する生体の応答機構を利用したモデル動物の作製を試みた.利用したのは,化学発癌の主因である親電子性物質に応答した生体防御遺伝子群の転写誘導機構である.これらの遺伝子の転写制御領域又はその制御を司る転写因子Nrf2の活性化機構の活用し,マウスとゼブラフィッシュを用いたモニター系構築を目指した.親電子性物質センサーからのシグナルは,Nrf2のN末に存在するNeh2ドメインに伝わり,何らかの機構によりNrf2の蛋白質安定化と核移行に促し,標的遺伝子の転写活性化に帰結する.Nrf2のN末半分にLacZ遺伝子を融合した構築をNrf2遺伝子座に挿入したマウス(Nrf2-LacZマウス)を作出したところ,親電子性物質を混ぜた餌の供与で通常検出できない小腸吸収上皮におけるLacZ発現が著名に誘導された.このことは,食物や医薬に混入する有害化学物質の検出に当該マウスが有用であることを示すとともに,Nrf2のN末に蛋白質安定性に関わる領域があることを指摘した.この領域をGFP融合蛋白質を利用して探索したところ,Neh2ドメインが浮かび上がった.一方,生体防御遺伝子であるNQO-1,GSTP,HO-1の転写制御領域を単離し,これにGFP遺伝子を融合した構築をもつトランスジェニックマウス及びゼブラフィッシュの系統化を試みた.一過性のレポーター解析により各構築の親電子性物質に対する応答能は確認できた.現在,ゲノムに同構築を安定にもつ系統の作製を試みている.計画遂行中の社会変化により,哺乳動物の大量殺戮は困難な情勢となってきたので,培養細胞を用いたモニター系の開発にも取り組むことにした.前述のNrf2-LacZマウスから繊維芽細胞を樹立したところ,親電子性試薬に対するLacZ誘導能を確認できた.現在,GFPを用いた系も立ち上げつつある.

研究成果

(31件)

すべてその他

すべて文献書誌 (31件)

[文献書誌] Enomoto, A.: "High sensitivity of Nrf2 knockout mice to acetoaminophen hepatotoxicity associated with decreased expression of ARE-regulated drug metabolizing enzyme and antioxidant genes"Toxicol. Sci.. 59. 169-177 (2001)
[文献書誌] Tabata, M.: "Stimulation of GATA-2 as a mechanism of hydrogen peroxide suppression in hypoxia-induced erythropoietin gene expression"J. Cell. Physiol.. 186. 260-267 (2001)
[文献書誌] Ramos-Gomez, M.: "Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice"Proc. Natl. Acad. Sci. USA. 98. 3410-3415 (2001)
[文献書誌] Kwak, M-K.: "Role of Transcription factor Nrf2 in the induction of hepatic phase 2 and antioxidative enzymes in vivo by the cancer chemopreventive agent, 3H-1, 2-dithiole-3-thione"Mol. Med.. 7. 135-145 (2001)
[文献書誌] Kanezaki, R.: "Transcription factor BACHI is recruited to nucleus by its novel alternative spliced isoform expressed in testis"J. Biol. Chem.. 276. 7278-7284 (2001)
[文献書誌] Kobayashi, M.: "The homeobox protein Six3 interacts with the Groucho corepressor and acts as a transcriptional repressor in eye and forebrain formation"Dev. Biol.. 232. 315-326 (2001)
[文献書誌] Hosoya, T.: "Defective development of secretary neurons in the hypothalamus of Arnt2-knockout mice"Genes Cells. 6. 361-374 (2001)
[文献書誌] Umetani, M.: "Function of GATA transcription factors in the induction of endothelial VCAM-1 by TNF-α"Arterioscler. Thromb. Vasc. Biol.. 21. 917-922 (2001)
[文献書誌] Kobayashi, M.: "Positive regulation by GATA-1 of GATA-1 gene hematopoietic regulatory domain in zebrafish embryos in vivo"Development. 128. 2341-2350 (2001)
[文献書誌] Kim, Y-C.: "Hemin-induced activation of the Thioredoxin gene by Nrf2: a differential regulation of the antioxidant responsive element(ARE) by switch of its binding factors"J. Biol. Chem.. 276. 18399-18406 (2001)
[文献書誌] Ogawa, K.: "Heme mediates de-repression of Maf recognition element through direct binding to transcription factor Bach1"EMBO J.. 20. 2835-2843 (2001)
[文献書誌] Yoh, K.: "Nrf2 deficient mouse: a novel model for lupus nephritis"Kidney Int.. 60. 1343-1353 (2001)
[文献書誌] Katoh, Y.: "Two domains of Nrf2 cooperatively bind CBP(CREB binding protein) and synergistically activate transcription"Genes Cells. 6. 857-868 (2001)
[文献書誌] Shimizu, R.: "In vivo requirements for functional GATA-1 domains during primitive and definitive erythropoiesis"EMBO J.. 20. 5250-5260 (2001)
[文献書誌] Sun, J.: "The promoter of mouse transcription repressor bach1 is regulated by Sp1 and transactivated by Bach1"J. Biochem. 130. 385-392 (2001)
[文献書誌] McMahon, M.: "The Cap "n" Collar basic leucine zipper transcription factor Nrf2 controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes"Cancer Res.. 61. 3299-3307 (2001)
[文献書誌] Imagawa, S.: "Levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea-hypopnea syndrome"Blood. 98. 1255-1257 (2001)
[文献書誌] Kwak, M.K.: "Role of phase 2 enzyme induction in chemoprotection by dithiolethiones"Mutat Res.. 481. 305-315 (2001)
[文献書誌] Yoh, K.: "Transgenic overexpression of MafK suppresses T cell proliferation and function in vivo"Genes Cells. 6. 1055-1066 (2001)
[文献書誌] Fujimoto, T.: "Stepwise divergence of primitive and definitive hematopoietic and endothelial cell lineages"Genes Cells. 6. 1113-1127 (2001)
[文献書誌] Aoki, Y.: "Accelerated DNA adducts formation in the lung of the Nrf2 knockout mouse exposed to diesel exhaust"Toxicol. Appl. Pharmacol. 173. 154-160 (2001)
[文献書誌] Cho, H.Y.: "Linkage analysis of susceptibility to hyperoxic lung injury: role of Nrf2 as a candidate gene"Am. J. Res. Cell Mol. Biol.. (in press).
[文献書誌] Morimitsu, Y.: "A sulforaphane analogue that potentially activates the Nrf2-dependent detoxification pathway"J. Biol. Chem.. (in press).
[文献書誌] Kirito, K.: "A functional role of Stat3 in in vivo megakaryopoiesis"Blood. (in press).
[文献書誌] Kusunoki, H.: "Solution structure of the DNA-binding domain of MafG"Nature Str. Biol.. (in press).
[文献書誌] Kwak, M.K.: "Regulation of the expression of Nrf2 transcription factor by cancer chemopreventive agents through antioxidant response element (ARE)-like sequences in its proximal promoter region"Mol. Cell. Biol.. (in press).
[文献書誌] Ueno, Y.: "Elimination of Pasteurella pneumotropica from a Contaminated Mouse Colony by Oral Administration of Enrofloxacin"Exp. Animals. (in press).
[文献書誌] 山本雅之編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)
[文献書誌] 勝岡史城, 山本雅之: "新臨床医のための分子医学シリーズ/造血幹細胞研究のいまと臨床応用「遺伝子操作マウスを用いた血液疾患モデルの作製」"羊土社. (2001)
[文献書誌] 中島修, 山本雅之: "Annual Review 血液 2002 「X染色体連鎖型鉄芽球性貧血モデルおよび鉄代謝異常関連遺伝子改変動物の分子生物学的解析」"中外医学社(印刷中).
[文献書誌] 鈴木教郎, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ「造血幹細胞の転写調節機構」"中外医学社(印刷中).

2001 年度 実績報告書

外来異物に対する信号伝達機構を利用した環境モニター動物の開発

研究代表者

山本 雅之 筑波大学, 基礎医学系, 教授 (50166823)

研究成果

[文献書誌] Enomoto, A.: "High sensitivity of Nrf2 knockout mice to acetoaminophen hepatotoxicity associated with decreased expression of ARE-regulated drug metabolizing enzyme and antioxidant genes"Toxicol. Sci.. 59. 169-177 (2001)

[文献書誌] Tabata, M.: "Stimulation of GATA-2 as a mechanism of hydrogen peroxide suppression in hypoxia-induced erythropoietin gene expression"J. Cell. Physiol.. 186. 260-267 (2001)

[文献書誌] Ramos-Gomez, M.: "Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice"Proc. Natl. Acad. Sci. USA. 98. 3410-3415 (2001)

[文献書誌] Kwak, M-K.: "Role of Transcription factor Nrf2 in the induction of hepatic phase 2 and antioxidative enzymes in vivo by the cancer chemopreventive agent, 3H-1, 2-dithiole-3-thione"Mol. Med.. 7. 135-145 (2001)

[文献書誌] Kanezaki, R.: "Transcription factor BACHI is recruited to nucleus by its novel alternative spliced isoform expressed in testis"J. Biol. Chem.. 276. 7278-7284 (2001)

[文献書誌] Kobayashi, M.: "The homeobox protein Six3 interacts with the Groucho corepressor and acts as a transcriptional repressor in eye and forebrain formation"Dev. Biol.. 232. 315-326 (2001)

[文献書誌] Hosoya, T.: "Defective development of secretary neurons in the hypothalamus of Arnt2-knockout mice"Genes Cells. 6. 361-374 (2001)

[文献書誌] Umetani, M.: "Function of GATA transcription factors in the induction of endothelial VCAM-1 by TNF-α"Arterioscler. Thromb. Vasc. Biol.. 21. 917-922 (2001)

[文献書誌] Kobayashi, M.: "Positive regulation by GATA-1 of GATA-1 gene hematopoietic regulatory domain in zebrafish embryos in vivo"Development. 128. 2341-2350 (2001)

[文献書誌] Kim, Y-C.: "Hemin-induced activation of the Thioredoxin gene by Nrf2: a differential regulation of the antioxidant responsive element(ARE) by switch of its binding factors"J. Biol. Chem.. 276. 18399-18406 (2001)

[文献書誌] Ogawa, K.: "Heme mediates de-repression of Maf recognition element through direct binding to transcription factor Bach1"EMBO J.. 20. 2835-2843 (2001)

[文献書誌] Yoh, K.: "Nrf2 deficient mouse: a novel model for lupus nephritis"Kidney Int.. 60. 1343-1353 (2001)

[文献書誌] Katoh, Y.: "Two domains of Nrf2 cooperatively bind CBP(CREB binding protein) and synergistically activate transcription"Genes Cells. 6. 857-868 (2001)

[文献書誌] Shimizu, R.: "In vivo requirements for functional GATA-1 domains during primitive and definitive erythropoiesis"EMBO J.. 20. 5250-5260 (2001)

[文献書誌] Sun, J.: "The promoter of mouse transcription repressor bach1 is regulated by Sp1 and transactivated by Bach1"J. Biochem. 130. 385-392 (2001)

[文献書誌] McMahon, M.: "The Cap "n" Collar basic leucine zipper transcription factor Nrf2 controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes"Cancer Res.. 61. 3299-3307 (2001)

[文献書誌] Imagawa, S.: "Levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea-hypopnea syndrome"Blood. 98. 1255-1257 (2001)

[文献書誌] Kwak, M.K.: "Role of phase 2 enzyme induction in chemoprotection by dithiolethiones"Mutat Res.. 481. 305-315 (2001)

[文献書誌] Yoh, K.: "Transgenic overexpression of MafK suppresses T cell proliferation and function in vivo"Genes Cells. 6. 1055-1066 (2001)

[文献書誌] Fujimoto, T.: "Stepwise divergence of primitive and definitive hematopoietic and endothelial cell lineages"Genes Cells. 6. 1113-1127 (2001)

[文献書誌] Aoki, Y.: "Accelerated DNA adducts formation in the lung of the Nrf2 knockout mouse exposed to diesel exhaust"Toxicol. Appl. Pharmacol. 173. 154-160 (2001)

[文献書誌] Cho, H.Y.: "Linkage analysis of susceptibility to hyperoxic lung injury: role of Nrf2 as a candidate gene"Am. J. Res. Cell Mol. Biol.. (in press).

[文献書誌] Morimitsu, Y.: "A sulforaphane analogue that potentially activates the Nrf2-dependent detoxification pathway"J. Biol. Chem.. (in press).

[文献書誌] Kirito, K.: "A functional role of Stat3 in in vivo megakaryopoiesis"Blood. (in press).

[文献書誌] Kusunoki, H.: "Solution structure of the DNA-binding domain of MafG"Nature Str. Biol.. (in press).

[文献書誌] Kwak, M.K.: "Regulation of the expression of Nrf2 transcription factor by cancer chemopreventive agents through antioxidant response element (ARE)-like sequences in its proximal promoter region"Mol. Cell. Biol.. (in press).

[文献書誌] Ueno, Y.: "Elimination of Pasteurella pneumotropica from a Contaminated Mouse Colony by Oral Administration of Enrofloxacin"Exp. Animals. (in press).

[文献書誌] 山本雅之 編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)

[文献書誌] 勝岡史城, 山本雅之: "新臨床医のための分子医学シリーズ/造血幹細胞研究のいまと臨床応用「遺伝子操作マウスを用いた血液疾患モデルの作製」"羊土社. (2001)

[文献書誌] 中島修, 山本雅之: "Annual Review 血液 2002 「X染色体連鎖型鉄芽球性貧血モデルおよび鉄代謝異常関連遺伝子改変動物の分子生物学的解析」"中外医学社(印刷中).

[文献書誌] 鈴木教郎, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ 「造血幹細胞の転写調節機構」"中外医学社(印刷中).

2001 年度実績報告書

山本雅之筑波大学, 基礎医学系, 教授 (50166823)

[文献書誌] 山本雅之編: "「遺伝子の構造と機能」シリーズ・バイオサイエンスの新世紀"日本生化学会編集,共立出版. 178 (2001)

[文献書誌] 鈴木教郎, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ「造血幹細胞の転写調節機構」"中外医学社(印刷中).