Reines D, Ghanouni P, Li Q Q, Mote J
Graduate Program in Biochemistry and Molecular Biology, Emory University School of Medicine, Atlanta, Georgia 30322.
J Biol Chem. 1992 Aug 5;267(22):15516-22.
Regulation of transcription elongation is an important mechanism in controlling eukaryotic gene expression. SII is an RNA polymerase II-binding protein that stimulates transcription elongation and also activates nascent transcript cleavage by RNA polymerase II in elongation complexes in vitro (Reines, D. (1992) J. Biol. Chem. 267, 3795-3800). Here we show that SII-dependent in vitro transcription through an arrest site in a human gene is preceded by nascent transcript cleavage. RNA cleavage appeared to be an obligatory step in the SII activation process. Recombinant SII activated cleavage while a truncated derivative lacking polymerase binding activity did not. Cleavage was not restricted to an elongation complex arrested at this particular site, showing that nascent RNA hydrolysis is a general property of RNA polymerase II elongation complexes. These data support a model whereby SII stimulates elongation via a ribonuclease activity of the elongation complex.
转录延伸的调控是控制真核基因表达的一个重要机制。SII是一种与RNA聚合酶II结合的蛋白质,它能刺激转录延伸,并且在体外的延伸复合物中还能激活RNA聚合酶II对新生转录本的切割(Reines, D. (1992) 《生物化学杂志》267, 3795 - 3800)。在此我们表明,在人基因中通过一个停滞位点进行的依赖SII的体外转录之前会发生新生转录本的切割。RNA切割似乎是SII激活过程中的一个必要步骤。重组SII激活了切割,而缺乏聚合酶结合活性的截短衍生物则没有。切割并不局限于在这个特定位点停滞的延伸复合物,这表明新生RNA水解是RNA聚合酶II延伸复合物的一个普遍特性。这些数据支持了一个模型,即SII通过延伸复合物的核糖核酸酶活性来刺激延伸。