Kataoka N, Hashimoto S, Shimura Y
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Biochem Biophys Res Commun. 1993 Jan 15;190(1):223-8. doi: 10.1006/bbrc.1993.1034.
We investigated in vitro splicing reaction using an artificially created mRNA precursor containing a single 5' splice site and tandemly duplicated 3' splice sites. We found that the 3' splice site proximal to the 5' splice site is predominantly used under the standard splicing conditions. However, when the preheated nuclear extract was employed, the intermediate in which the distal 3' splice site was selected accumulated exclusively. This shows that heat treatment of nuclear extract abolishes the activity involved in the selection of the 3' splice sites that are in cis-competition for the common 5' splice site. The results presented here suggest the presence of a factor(s) required for the selection of the proximal 3' splice site.
我们使用人工构建的包含单个5'剪接位点和串联重复3'剪接位点的mRNA前体研究了体外剪接反应。我们发现，在标准剪接条件下，靠近5'剪接位点的3'剪接位点被优先使用。然而，当使用预热的核提取物时，仅积累了选择远端3'剪接位点的中间体。这表明核提取物的热处理消除了参与选择与共同5'剪接位点顺式竞争的3'剪接位点的活性。此处呈现的结果表明存在选择近端3'剪接位点所需的一种或多种因子。
