2001 Fiscal Year Annual Research Report

クロマチン変換による遺伝子転写の標的型操作技術の開発

Research Project

Project/Area Number	13557015
Research Institution	Kumamoto University
Principal Investigator	中尾光善熊本大学, 発生医学研究センター, 教授 (00217663)
Co-Investigator(Kenkyū-buntansha)	谷上信大塚GEN研究所, 所長
Keywords	エピジェネティクス / DNAメチル化 / クロマチン / メチル化CpG結合タンパク質 / 転写 / 遺伝子治療
Research Abstract	ヒトの疾患の分子機構を考察すると、病因遺伝子の異常な活性化または不活性化が病態に直接関わる例が多数知られている。また、遺伝子治療ベクターなどが宿主細胞内でしばしばメチル化を受けて不活性化される。細胞内の遺伝子発現を選択的に人為的に操作できる技術として、クロマチン変換による新しい標的型遺伝子操作技術の開発を推進している。具体的には、[1]メチル化CpG結合タンパク質の転写抑制ドメインを用いた転写抑制技術、[2]DNAメチル化酵素の活性領域を用いた恒久的な転写抑制技術、[3]変異型メチル化DNA結合ドメインを用いたメチル化遺伝子の活性化技術に取り組んだ。[1]発がんを促進する転写因子E2F1のDNA結合ドメインとMBD1よびMeCP2の転写抑制ドメインを連結したキメラ蛋白質を作成して、標的型の転写抑制技術を開発した。E2F1の標的遺伝子であるサイクリンDのプロモーター活性を低下させた。[2]転写因子のDNA結合ドメインとDNAメチル化酵素活性領域を連結させた標的型メチル化酵素の開発に取り組んでいる。標的遺伝子が新規にメチル化されること、獲得されたメチル化が体細胞分裂で安定に維持されることを期待して、がん細胞の増殖を阻止を目標にしている。いずれも、プラスミドおよびウイルスベクターを用いて、幅広い転写操作技術としての応用を目指す予定である。[3]メチル化DNA結合ドメインの変異体がドミナントネガティブ効果を持つことを見い出したので、メチル化プロモーターを再活性化する技術にも取り組む。

Research Products

(12 results)

All Other

All Publications (12 results)

[Publications] Ohki, I.: "Solution structure of the methyl-CpG-binding domain of human MBD1 in complex with a methylated DNA"Cell. 105. 487-497 (2001)
[Publications] Takizawa, T.: "DNA methylation is a critical cell-intrinsic determinant of astrocyte differentiation in the fetal brain"Dev. Cell. 1. 749-758 (2001)
[Publications] Meguro, M.: "A novel maternally expressed gene, ATP10C, encodes a putative aminophospholipid translocase associated with Angelman syndrome"Nature Genet.. 28. 19-20 (2001)
[Publications] Nakao, M.: "Epigenetics : interactions of DNA methylation and chromatin"Gene. 278. 25-31 (2001)
[Publications] Honda, Y.: "Cooperation of HECT-domain ubiquitin ligase hHYD and DNA topoisomerase II-binding protein for DNA damage response"J. Biol. Chem.. (in press).
[Publications] EL-Gazzar, M.A.: "PU. 1 is involved in the regulation of B lineage-associated and developmental stage-dependent expression of the germinal center-associated DNA-primase GANP"J. Biol. Chem.. 276. 48000-48008 (2001)
[Publications] Kino, T.: "Identification of the cis-acting region in the NF2 gene promoter as a potential target for mutation and methylation-dependent silencing in schwannoma"Genes Cells. 6. 441-454 (2001)
[Publications] Takebayashi, S.: "5-aza-2'-deoxycytidine induces histone hyperacetylation of mouse centromeric heterochromatin by a mechanism independent of DNA demethylation"Biochem. Biophys. Res. Commun. 288. 921-926 (2001)
[Publications] Meguro, M.: "Large-scale evaluation of imprinting status in the Prader-Willi syndrome region : an imprinted direct repeat cluster resembling small nucleolar RNA genes"Hum. Mol. Genet.. 10. 83-394 (2001)
[Publications] Sudo, T.: "Activation of Cdh1-dependent APC is required for G1 cell cycle arrest and DNA damage-induced G2 checkpoint in vertebrate cells"EMBO J.. 20. 6499-6508 (2001)
[Publications] Kaneko, T.: "Calpain-dependent proteolysis of merlin occurs by oxidative stress in meningiomas"Cancer. 92. 2662-2672 (2001)
[Publications] Nakao, M.: "Regulation of transcription and chromatin by methyl-CpG binding protein MBD1"Brain Dev.. 23 Suppl 1. S174-S176 (2001)

2001 Fiscal Year Annual Research Report

クロマチン変換による遺伝子転写の標的型操作技術の開発

Principal Investigator

中尾 光善 熊本大学, 発生医学研究センター, 教授 (00217663)

Research Products

[Publications] Ohki, I.: "Solution structure of the methyl-CpG-binding domain of human MBD1 in complex with a methylated DNA"Cell. 105. 487-497 (2001)

[Publications] Takizawa, T.: "DNA methylation is a critical cell-intrinsic determinant of astrocyte differentiation in the fetal brain"Dev. Cell. 1. 749-758 (2001)

[Publications] Meguro, M.: "A novel maternally expressed gene, ATP10C, encodes a putative aminophospholipid translocase associated with Angelman syndrome"Nature Genet.. 28. 19-20 (2001)

[Publications] Nakao, M.: "Epigenetics : interactions of DNA methylation and chromatin"Gene. 278. 25-31 (2001)

[Publications] Honda, Y.: "Cooperation of HECT-domain ubiquitin ligase hHYD and DNA topoisomerase II-binding protein for DNA damage response"J. Biol. Chem.. (in press).

[Publications] EL-Gazzar, M.A.: "PU. 1 is involved in the regulation of B lineage-associated and developmental stage-dependent expression of the germinal center-associated DNA-primase GANP"J. Biol. Chem.. 276. 48000-48008 (2001)

[Publications] Kino, T.: "Identification of the cis-acting region in the NF2 gene promoter as a potential target for mutation and methylation-dependent silencing in schwannoma"Genes Cells. 6. 441-454 (2001)

[Publications] Takebayashi, S.: "5-aza-2'-deoxycytidine induces histone hyperacetylation of mouse centromeric heterochromatin by a mechanism independent of DNA demethylation"Biochem. Biophys. Res. Commun. 288. 921-926 (2001)

[Publications] Meguro, M.: "Large-scale evaluation of imprinting status in the Prader-Willi syndrome region : an imprinted direct repeat cluster resembling small nucleolar RNA genes"Hum. Mol. Genet.. 10. 83-394 (2001)

[Publications] Sudo, T.: "Activation of Cdh1-dependent APC is required for G1 cell cycle arrest and DNA damage-induced G2 checkpoint in vertebrate cells"EMBO J.. 20. 6499-6508 (2001)

[Publications] Kaneko, T.: "Calpain-dependent proteolysis of merlin occurs by oxidative stress in meningiomas"Cancer. 92. 2662-2672 (2001)

[Publications] Nakao, M.: "Regulation of transcription and chromatin by methyl-CpG binding protein MBD1"Brain Dev.. 23 Suppl 1. S174-S176 (2001)

中尾光善熊本大学, 発生医学研究センター, 教授 (00217663)