人类拼接因子ASF的初级结构揭示了与Drosophila调节器的相似之处
H Ge1, P Zuo, J L Manley
1Department of Biological Sciences, Columbia University, New York, New York 10027.
Cell
|July 26, 1991
概括
研究人员分离了ASF-1cDNA,编码了替代拼接因子 (ASF) 蛋白质,它调节了基因拼接. 再组合的ASF蛋白激活了splicing和替代的5' splice-site选择在体外.
科学领域:
- 分子生物学分子生物学
- RNA剪接机制的机制
- 基因规则 基因规则
背景情况:
- 替代拼接因子 (ASF) 是一种先前纯化的人类蛋白质.
- 在SV40早期前mRNA中,ASF调节了替代的5'拼接部位利用.
研究的目的:
- 隔离编码ASF蛋白的cDNA,被指定为ASF-1.
- 描述ASF-1蛋白及其在拼接中的功能.
- 调查ASF前mRNA的替代拼接方法.
主要方法:
- cDNA的分离和测序. cDNA的分离和测序.
- 重组蛋白表达 (大肠杆菌) 和体外拼接试验.
- 分析额外的cDNA来研究ASF前mRNA拼接.
主要成果:
- 隔离的ASF-1cDNA编码248氨基酸蛋白质,具有RNA结合域和富含氨酸/氨酸的C端.
- 再组合的ASF-1激活了splicing和替代的5'splice位置选择在体外.
- ASF前mRNA经历了替代拼接,生成ASF-2mRNA,并转化为替代的读取框架.
结论:
- 隔离的ASF-1cDNA编码了一个功能性的替代拼接因子.
- 再组合的ASF蛋白质足以进行拼接激活和拼接位置切换.
- ASF前mRNA的替代拼接产生了不同的mRNA异型,这对蛋白质表达有影响.
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