SITE-SPECIFIC MODIFICATION OF PRE-MESSENGER-RNA - THE 2'-HYDROXYL GROUPS AT THE SPLICE SITES
SITE-SPECIFIC MODIFICATION OF PRE-MESSENGER-RNA - THE 2'-HYDROXYL GROUPS AT THE SPLICE SITES
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DOI：
10.1126/science.1589782
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发表时间：
1992-05-15
期刊：
影响因子：
56.9
通讯作者：
SHARP, PA
中科院分区：
文献类型：
--
作者：
MOORE, MJ;SHARP, PA
开发了一种简单有效的合成长的、位点特异性修饰的RNA分子的方法，其中使用噬菌体T4 DNA连接酶连接RNA片段。在核前信使RNA底物的任一剪接位点，用单个氢或O-甲基取代2 ′-羟基。修饰的前信使RNA的体外剪接揭示，尽管在任一剪接位点处2 '-羟基不是绝对需要的，但在3'剪接位点处的2 '-羟基对于剪接的第二步是重要的。这些结果进行了比较，在自我剪接四膜虫组I内含子类似的2 '-羟基基团的以前的研究。
A simple and efficient method for synthesizing long, site-specifically modified RNA molecules was developed whereby segments of RNA were joined with the use of bacteriophage T4 DNA ligase. A single hydrogen or O-methyl group was substituted for the 2'-hydroxyl group at either splice site of a nuclear pre-messenger RNA substrate. Splicing of the modified pre-messenger RNA's in vitro revealed that, although a 2'-hydroxyl is not absolutely required at either splice site, the 2'-hydroxyl at the 3' splice site is important for the second step of splicing. These results are compared to previous studies of analogous 2'-hydroxyl groups in the self-splicing Tetrahymena group I intron.