The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction.
The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction.
复制标题
用补骨脂素探测的信使 RNA 前体的二级结构在体外剪接反应中被熔化。
DOI：
10.1093/nar/15.22.9279
复制
发表时间：
1987
影响因子：
14.9
通讯作者：
Goldenberg,CJ
中科院分区：
文献类型：
--
作者：
Wollenzien,PL;Goswami,P;Teare,J;Szeberenyi,J;Goldenberg,CJ
SP6/小鼠胰岛素前体RNA的二级结构通过补骨脂素交联实验测定。一系列长程接触发生在包含插入序列的前 mRNA 的左半部分内。由于一些相互作用共享共同位点，因此前 mRNA 存在多个二级结构。在剪接缓冲液中但不添加剪接提取物时，许多这些相互作用在至少 50°C 的温度下保持稳定。然而，这些相互作用在体外剪接反应期间被解离。这种解离需要 ATP，并且发生在剪接反应的前 30 分钟内。纯化 RNA 分子内不同位置处含有补骨脂素交联的前 mRNA，并用作体外剪接的底物。任何双链区域的补骨脂素交联都会导致剪接反应的完全抑制。这表明 SP6/小鼠胰岛素前体 mRNA 二级结构的不稳定对于体外剪接是必要的。
The secondary structure of the SP6/mouse insulin precursor RNA was determined by psoralen cross-linking experiments. A series of long-range contacts occur within the left half of the pre-mRNA that contains the intervening sequence. Multiple secondary structures for the pre-mRNA exist since some of the interactions share common sites. In splicing buffer but without the splicing extract added, many of these interactions are stable up to at least 50°C. These interactions, however, are dissociated during thein vitrosplicing reaction. This dissociation requires ATP and it occurs during the first 30 min, of the splicing reaction. Pre-mRNAs containing psoralen cross-links in different locations within the RNA molecule were purified and used as substrates forin vitrosplicing. Psoralen cross-links at any of the double-stranded regions resulted in complete inhibition of the splicing reaction. This indicates that destabilization of the secondary structure of the SP6/mouse insulin pre-mRNA is necessary forin vitrosplicing.