Price D H, Parker C S
Cell. 1984 Sep;38(2):423-9. doi: 10.1016/0092-8674(84)90497-5.
We have examined the process by which the 3' terminus of the Drosophila histone H3 mRNA is produced in vitro. When a template containing a portion of DNA that flanks the normal 3' end of the histone H3 gene and an oligo dC tail on the template strand is transcribed in vitro by Drosophila RNA polymerase II, transcription continues beyond the 3' end of the H3 gene. A processing activity was identified that cleaves the precursor transcript generating an RNA species with the same 3' end as the mature H3 mRNA. The processing activity was partially purified by ion exchange chromatography and sucrose gradient sedimentation. The isolated activity was found to require Mg++ but did not require addition of a nucleoside triphosphate for activity. The activity sedimented with a molecular weight of approximately 140,000 daltons. Transcription of the template and processing of the RNA can be uncoupled in vitro.
我们研究了果蝇组蛋白H3 mRNA 3'末端在体外产生的过程。当用果蝇RNA聚合酶II在体外转录一个模板时,该模板包含组蛋白H3基因正常3'端侧翼的一段DNA以及模板链上的一个寡聚dC尾,转录会持续到H3基因3'端之外。鉴定出一种加工活性,它切割前体转录本,产生一种与成熟H3 mRNA具有相同3'端的RNA分子。通过离子交换色谱和蔗糖梯度沉降对该加工活性进行了部分纯化。发现分离出的活性需要Mg++,但活性不需要添加核苷三磷酸。该活性沉降时的分子量约为140,000道尔顿。模板的转录和RNA的加工在体外可以解偶联。