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从培养的大鼠神经细胞中克隆一种新型甲状腺/类固醇受体超家族基因。

Molecular cloning of a novel thyroid/steroid receptor superfamily gene from cultured rat neuronal cells.

作者信息

Ohkura N, Hijikuro M, Yamamoto A, Miki K

机构信息

Terumo R&D Center, Kanagawa, Japan.

出版信息

Biochem Biophys Res Commun. 1994 Dec 30;205(3):1959-65. doi: 10.1006/bbrc.1994.2900.

Abstract

Brain neuronal cells in primary culture provide one system for the analysis of neuronal development, including apoptosis. By PCR amplification, with primers designed from conserved sequences among nuclear receptors, a novel member of the thyroid/steroid receptor superfamily was identified from forebrain neuronal cells undergoing apoptosis and designated as NOR-1 (neuron derived orphan receptor). A cDNA clone of NOR-1 encodes a 68kDa, 628 amino acid residue-containing protein, with an amino acid sequence highly homologous to the Nur77 family in its DNA-binding domain and moderately so in its putative ligand-binding domain. Using mobility shift assay, we showed that the NOR-1 protein binds to the B1a response-element which has been identified as the target sequence of the Nur77 family, suggesting that three members of the Nur77 family may transactivate common target gene(s) at different situations.

摘要

原代培养的脑神经元细胞为分析神经元发育(包括细胞凋亡)提供了一个系统。通过PCR扩增,利用从核受体保守序列设计的引物,从正在经历凋亡的前脑神经元细胞中鉴定出甲状腺/类固醇受体超家族的一个新成员,并将其命名为NOR-1(神经元衍生孤儿受体)。NOR-1的cDNA克隆编码一个含628个氨基酸残基的68kDa蛋白,其氨基酸序列在DNA结合结构域与Nur77家族高度同源,在假定的配体结合结构域中度同源。使用迁移率变动分析,我们表明NOR-1蛋白与已被确定为Nur77家族靶序列的B1a反应元件结合,这表明Nur77家族的三个成员可能在不同情况下反式激活共同的靶基因。

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