Reeder T C, Hawley D K
Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene 97403, USA.
Cell. 1996 Nov 15;87(4):767-77. doi: 10.1016/s0092-8674(00)81395-1.
The adenovirus major late arrest site blocks transcription by mammalian RNA polymerase II in vitro downstream of the major late promoter but not the mouse beta-globin promoter. We localized the sequences responsible for anti-arrest to the 5' end of the beta-globin transcript and demonstrated that anti-arrest required that this region of RNA form base pairs with the nascent transcript upstream of the arrest site. Small antisense RNA or DNA oligonucleotides hybridizing upstream of the arrest site also prevented arrest when added in trans. Our results suggest that arrest is accompanied by retraction of the nascent transcript into the interior of the polymerase and that hybridization of the transcript prevents this movement, thereby allowing the polymerase to continue elongation.
腺病毒主要晚期终止位点在体外可阻断哺乳动物RNA聚合酶II在主要晚期启动子下游的转录,但不影响小鼠β-珠蛋白启动子。我们将负责抗终止的序列定位到β-珠蛋白转录本的5'端,并证明抗终止需要RNA的这一区域与终止位点上游的新生转录本形成碱基对。当反式添加与终止位点上游杂交的小反义RNA或DNA寡核苷酸时,也能防止终止。我们的结果表明,终止伴随着新生转录本缩回到聚合酶内部,而转录本的杂交可阻止这种移动,从而使聚合酶能够继续延伸。