哺乳动物拼接因子U2AF的克隆和域结构
P D Zamore1, J G Patton, M R Green
1Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Nature
|February 13, 1992
概括
研究人员为U2 snRNP辅助因子65 (U2AF65) 大子单元分离并表达了一个互补的DNA克隆. 这种必不可少的拼接因子含有不同的RNA结合和拼接特定的域.
科学领域:
- 分子生物学分子生物学
- RNA拼接机制的机制
- 基因表达规范 基因表达规范
背景情况:
- 前传递 RNA (mRNA) 拼接是基因表达的一个关键过程.
- U2 snRNP辅助因子 (U2AF) 对于哺乳动物mRNA拼接至关重要.
- U2AF包括一个大子单位 (U2AF65) 和一个小子单位.
研究的目的:
- 隔离和表达编码U2AF (U2AF65) 大子单元的互补DNA (cDNA).
- 为了描述U2AF65蛋白的功能域.
- 了解U2AF65在mRNA前拼接中的作用的分子基础.
主要方法:
- 补充DNA (cDNA) 克隆和分离.
- 在体外表达U2AF65蛋白.
- 对U2AF65.65的功能域分析.
主要成果:
- 一个编码U2AF65大亚单元的cDNA克隆被成功分离并在体外表达.
- U2AF65蛋白含有两个不同的功能域.
- 一个域通过核核蛋白共识序列 (RNP-CS) 域表现出序列特定的RNA结合.
- 另一个域的特点是氨酸/氨酸丰富的基因对剪接至关重要,但不直接与mRNA前结合.
结论:
- 隔离的U2AF65cDNA编码了一个具有特征域的功能蛋白.
- 该RNP-CS域调解序列特定的RNA结合.
- 富含氨酸/氨酸的图案对于U2AF65.5的拼接活动至关重要.
- 这些发现为U2AF65在mRNA前拼接中的分子机制提供了洞察力.
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