• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Multiple splicing factors are released from endogenous complexes during in vitro pre-mRNA splicing.
【24h】

Multiple splicing factors are released from endogenous complexes during in vitro pre-mRNA splicing.

机译：在体外预mRNA剪接过程中，多种剪接因子从内源性复合物中释放出来。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Pre-mRNA splicing occurs in a macromolecular complex called the spliceosome. Efforts to isolate spliceosomes from in vitro splicing reactions have been hampered by the presence of endogenous complexes that copurify with de novo spliceosomes formed on added pre-mRNA. We have found that removal of these large complexes from nuclear extracts prevents the splicing of exogenously added pre-mRNA. We therefore examined these complexes for the presence of splicing factors and proteins known or thought to be involved in RNA splicing. These fast-sedimenting structures were found to contain multiple small nuclear ribonucleoproteins (snRNPs) and a fragmented heterogeneous nuclear ribonucleoprotein complex. At least two splicing factors other than the snRNPs were also associated with these large structures. Upon incubation with ATP, these splicing factors as well as U1 and U2 snRNPs were released from these complexes. The presence of multiple splicing factors suggests that these complexes may be endogenous spliceosomes released from nuclei during preparation of splicing extracts. The removal of these structures from extracts that had been preincubated with ATP yielded a splicing extract devoid of large structures. This extract should prove useful in the fractionation of splicing factors and the isolation of native spliceosomes formed on exogenously added pre-mRNA.
机译：mRNA前剪接发生在称为剪接体的大分子复合物中。从体内剪接反应中分离剪接体的努力已经受到与在添加的前mRNA上形成的从头剪接体共纯化的内源性复合物的存在的阻碍。我们已经发现，从核提取物中除去这些大的复合物可防止外源添加的pre-mRNA的剪接。因此，我们检查了这些复合物是否存在剪接因子和已知或认为与RNA剪接有关的蛋白质。发现这些快速沉淀的结构包含多个小核核糖核蛋白（snRNPs）和一个片段化的异质核核糖核蛋白复合物。除了snRNPs以外，至少还有两个剪接因子也与这些大结构有关。与ATP孵育后，这些剪接因子以及U1和U2 snRNPs从这些复合物中释放出来。多种剪接因子的存在表明这些复合物可能是剪接提取物制备过程中从细胞核释放的内源剪接体。从已经与ATP预温育的提取物中除去这些结构，得到的剪接提取物没有大的结构。该提取物在剪接因子的分离以及在外源添加的pre-mRNA上形成的天然剪接体的分离中应被证明是有用的。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号