White R A, Kunkel G R
Department of Biochemistry and Biophysics, Texas A & M University, College Station 77843-2128.
Biochem Biophys Res Commun. 1993 Sep 30;195(3):1394-400. doi: 10.1006/bbrc.1993.2198.
In order to explore the coupling of transcription with splicing in mammalian cells we have prepared hybrid genes in which either the human U2 promoter, recognized by RNA polymerase II, or the human U6 promoter, recognized by RNA polymerase III, was fused to an intron-containing gene segment. Neither human small nuclear RNA gene contains an intron although U6 genes from some species of yeast contain a short intervening sequence. Following transfection of human cells and analysis of specific RNAs by primer extension we found that the chimeric U2 promoter-derived transcript was efficiently spliced but the RNA polymerase III transcript driven by the U6 promoter remained unspliced. Hence, the splicing apparatus differentiates between transcripts produced from two closely related promoters that are distinguished by RNA polymerase selectivity.
为了探究哺乳动物细胞中转录与剪接的偶联,我们制备了杂种基因,其中由RNA聚合酶II识别的人U2启动子或由RNA聚合酶III识别的人U6启动子与含内含子的基因片段融合。尽管来自某些酵母物种的U6基因含有短的间隔序列,但人小核RNA基因均不包含内含子。在转染人细胞并用引物延伸分析特定RNA后,我们发现嵌合的U2启动子衍生的转录本被有效剪接,但由U6启动子驱动的RNA聚合酶III转录本仍未剪接。因此,剪接装置能够区分由两个密切相关的启动子产生的转录本,这两个启动子通过RNA聚合酶选择性来区分。