3′-Box-dependent processing of human pre-U1 snRNA requires a combination of RNA and protein co-factors
3′-Box-dependent processing of human pre-U1 snRNA requires a combination of RNA and protein co-factors
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DOI：
10.1093/nar/gkh619
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发表时间：
2004-06-01
影响因子：
14.9
通讯作者：
Murphy, S
中科院分区：
文献类型：
--
作者：
Uguen, P;Murphy, S
通过体外系统，我们最近发现，由RNA聚合酶II合成的人类前snRNA的3‘端是由snRNA基因特异性3’盒指导的RNA加工产生的。为了完整地表征这一加工反应，我们进一步研究了pre-U1 snRNA正确形成3'端的体外要求。在这里，我们表明5'帽起刺激作用，加工需要磷酸肌酸。我们的结果还表明，u1预处理活动是热敏的，并且需要RNA成分。此外，与3‘ box相邻的确切序列影响体外产生的pre-U1 3’端的位置。有趣的是，对3‘-box依赖性加工具有活性的加工提取物也含有一种活性，该活性可将含有U1 Sm蛋白结合位点和3’末端茎环的RNA的3'端转化为成熟形式。
Using an in vitro system we have recently shown that the 3' ends of human pre-snRNAs synthesized by RNA polymerase II are produced by RNA processing directed by the snRNA gene-specific 3' box. Towards a complete characterization of this processing reaction we have further investigated the in vitro requirements for proper 3' end formation of pre-U1 snRNA. Here we show that the 5' cap plays a stimulatory role and processing requires creatine phosphate. Our results also indicate that the pre-U1 processing activity is heat sensitive and that an RNA component is required. In addition, the exact sequence adjacent to the 3' box influences the position of the pre-U1 3' end produced in vitro. Interestingly, the processing extract active for 3'-box-dependent processing also contains an activity that converts the 3' end of RNA containing the U1 Sm protein binding site and the 3' terminal stem-loop into the mature form.