一个编码具有具有E2F类属性的视网膜母细胞瘤结合蛋白的cDNA的表达克隆
W G Kaelin1, W Krek, W R Sellers
1Dana Farber Cancer Institute, Boston, Massachusetts.
Cell
|August 3, 1992
概括
研究人员确定了RBAP-1,一种与视网膜母细胞瘤 (RB) 蛋白口袋结合的蛋白质. RBAP-1 是E2F的一种形式,这是一个转录因子,对细胞生长控制至关重要.
科学领域:
- 分子生物学分子生物学
- 癌症研究 癌症研究
- 病毒学 病毒学
背景情况:
- 视网膜母细胞瘤 (RB) 蛋白质是细胞生长的关键调节者.
- RB蛋白的"口袋"域与细胞转录因子 (如E2F) 相互作用.
- E2F与控制细胞增殖有关.
研究的目的:
- 为了确定与RB蛋白相互作用的新型蛋白质.
- 描述一种新发现的RB结合蛋白的功能和结合特性.
主要方法:
- 构建和使用一种谷氨S转移酶 (GST) -视网膜母细胞瘤 (RB) 融合蛋白表达系统,用于快速蛋白质合成.
- 对lambda gt11表达库进行选,以识别RB结合蛋白.
- 共同净化试验,蛋白质与蛋白质相互作用研究和DNA结合试验,以表征RBAP-1.
主要成果:
- 克隆了编码RBAP-1的cDNA,它直接与RB口袋结合.
- 发现RBAP-1与E2F共同净化,并与腺病毒E4ORF 6/7蛋白特异相互作用.
- RBAP-1直接与E2FDNA识别序列结合,并具有功能性事务激活域.
结论:
- RBAP-1被确定为E2F的一种物种.
- RBAP-1 特别与RB口袋结合,这表明它在RB介导的细胞生长控制中的作用.
- 这些发现阐明了RB/E2F通路中的新型相互作用及其潜在的病毒蛋白调节.
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