人类增强因子TEF-1的克隆,表达和转录性质
J H Xiao1, I Davidson, H Matthes
1Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Génie Génétique et de Biologie, Faculté de Médecine, Strasbourg, France.
Cell
|May 17, 1991
概括
SV40转录增强因子1 (TEF-1) 的翻译始于AU编码子. 克隆的TEF-1结合了特定的DNA元素并抑制了活性,这表明细胞特异性的限制因子介导了其转激活功能.
科学领域:
- 分子生物学分子生物学
- 基因规则 基因规则
- 转录因子 转录因子
背景情况:
- SV40转录增强因子1 (TEF-1) 是基因表达的关键调节者.
- 对于TEF-1活动的精确机制,特别是其细胞特异性调节,目前尚不完全理解.
研究的目的:
- 为了描述编码TEF-1的cDNA.
- 调查TEF-1的翻译启动地点.
- 阐明TEF-1在不同细胞类型中的转录活性和调节机制.
主要方法:
- cDNA克隆和TEF-1的测序.
- 在体内和体内翻译试验.
- 使用SV40增强子 (GT-IIC和Sph) 的DNA结合试验.
- 在HeLa和MPC11细胞中进行记者基因测试.
- 构建和测试GAL4-TEF-1仿制蛋白.
主要成果:
- 克隆了TEF-1cDNA,并且证明翻译在体内仅在AUU编码中启动.
- 克隆的TEF-1特别与SV40 GT-IIC和Sph增强器结合.
- 在淋巴细胞MPC11细胞中,增强子是不活跃的,克隆的TEF-1抑制了内生TEF-1活动.
- 镇压是通过干扰/压制进行的,这是GAL4-TEF-1仿真体所证明的.
- 化学蛋白刺激HeLa细胞的转录,但不是MPC11细胞,表明细胞特异性因素.
结论:
- TEF-1翻译启动是特定的,发生在一个AUU编码子.
- TEF-1的转激活功能很可能是由一个高度限制的,可能是细胞特异的,可定位的转录中间因子介导的.
- 观察到的抑制和细胞特异性活动凸显了控制TEF-1功能的复杂调节机制.
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