Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G2W1.
Biochem Biophys Res Commun. 2010 Mar 12;393(3):461-5. doi: 10.1016/j.bbrc.2010.02.023. Epub 2010 Feb 10.
The maturation of many small nuclear RNAs is dependent on RNase III-like endonuclease mediated cleavage, which generates a loading site for the exosome complex that trims the precursor at its 3' end. Using a temperature sensitive Pac1 nuclease, here we show that the endonuclease cleavage is equally important in terminating the transcription of the U2 snRNA in Schizosaccharomyces pombe. Using a temperature sensitive Dhp1p 5'-->3' exonuclease, we demonstrate that it also is an essential component of the termination pathway. Taken together the results support a "reversed torpedoes" model for the termination and maturation of the U2 snRNA; the Pac1 endonuclease cleavage provides entry sites for the 3' and 5' exonuclease activities, leading to RNA maturation in one direction and transcript termination in the other.
许多小核 RNA 的成熟依赖于 RNase III 样内切酶介导的切割,该切割生成了一个加载位点,供外切体复合物在其 3' 末端修剪前体。在这里,我们使用温度敏感的 Pac1 核酸酶表明,内切酶切割在终止酿酒酵母中的 U2 snRNA 的转录中同样重要。使用温度敏感的 Dhp1p 5'-->3' 外切酶,我们证明它也是终止途径的必需组成部分。总之,这些结果支持 U2 snRNA 终止和成熟的“反向鱼雷”模型;Pac1 内切酶切割为 3' 和 5' 外切酶活性提供了进入位点,导致 RNA 在一个方向上成熟,而在另一个方向上终止转录。