Establishment of nuclear receptor-mediated assay for sex-steroid like compounds

核受体介导的性类固醇样化合物测定方法的建立

• 批准号: 11556065
• 负责人: KATO Shigeaki
• 金额: $ 8.32万
• 依托单位: Institute of Molecular and Cellular Biosciences, The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11556065/
• 关键词: sex steroid recepter; cofactors; in vitro interaction; transactivation; hormone-like compound; yeast two-hybrid assay; receptor cofactor; cDNA screening; ホルモン関連化合物; ステロイドホルモンレセプター; 共役転写因子群; 転写促進領域; ERα・ERβ; AR; クロマチン構造

Steroid hormones exert their actons through cognate nuclear receptors, which act ligand-inducible transcriptional factors. For this ligand-induced transactivation, ligand-bound nuclear receptors, nuclear receptors depend on ligand binding, but it is induced only by agonist, but not antgonist. In the present study, we searched novel coactivators for nuclear receptors for estrogen, androgen mineralcorticoid, vitamin D by biochemical approaches and yeast two-hybrid system, and found several candidate cDNAs. Especially, we found that p68/p72 acts as a co-activator specific for ERα. Moreover, we developed an assay system based on the interactions of coactivators with nuclear receptors in yeast to evaluate agonistic/antagonistid activity of related compounds.

类固醇激素通过同源核受体发挥作用，这些受体作用于配体诱导的转录因子。对于这种配体诱导的反式激活，配体结合核受体，核受体依赖于配体结合，但它只被激动剂诱导，而不是拮抗剂。本研究利用生物化学方法和酵母双杂交系统，寻找雌激素、雄激素、盐皮质激素、维生素D核受体的新辅激活因子，并筛选出了几个候选cDNA。特别是，我们发现p68/p72作为ERα特异性的共激活剂。此外，我们开发了一种基于辅激活因子与酵母核受体相互作用的测定系统，以评估相关化合物的激动/拮抗活性。

项目成果

Tai,H.,Kubota,N.,Kato,S.: "Involvement of nuclear receptor coactivator SRC-1 in estrogen-dependent cell growth of MCF-7 cells"Biochem.Biophys.Res.Commun.. in press. (2000)

Tai,H.,Kubota,N.,Kato,S.：“核受体辅激活剂 SRC-1 在 MCF-7 细胞雌激素依赖性细胞生长中的参与”Biochem.Biophys.Res.Commun.. 正在出版。

Kodera,Y.,Takeyama,K.,Kato,S., et al.: "Ligand-type specific interactions of peroxisome proliferator- activated receptor gamma with transcriptional coactivators"J.Biol.Chem.. 275. 33201-33204 (2000)

Kodera，Y.，Takeyama，K.，Kato，S.，等：“过氧化物酶体增殖物激活受体γ与转录共激活剂的配体型特异性相互作用”J.Biol.Chem.. 275. 33201-33204 (2000

Takeyama,K.,Kato.S.,et al.: "Selective interaction of vitamin D receptor with transcriptional coactivators by a vitamin D analog"Mol.Cell.Biol.. 19. 1409-1055 (1999)

Takeyama,K.,Kato.S.等人：“维生素 D 类似物对维生素 D 受体与转录共激活剂的选择性相互作用”Mol.Cell.Biol.. 19. 1409-1055 (1999)

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

Sekine,K.、Kato,S. 等人：“FGF10 对于肢体和肺的形成至关重要”《Nature Genetics》。

Li,M.,Indra,A.,Kato,S., et al.: "Skin abnormalities generated by temporally-controlled RXRα mutations in adult mouse epidermis."Nature. 407. 633-636 (2000)

Li, M.、Indra, A.、Kato, S. 等人：“成年小鼠表皮中时间控制的 RXRα 突变产生的皮肤异常。”《自然》，407. 633-636 (2000)