RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes.
RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes.
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由 HeLa 细胞分离组分形成的 RNA 剪接产物与快速沉降的复合物相关。
DOI：
10.1073/pnas.83.4.887
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发表时间：
1986
影响因子：
11.1
通讯作者：
Hurwitz,J
中科院分区：
文献类型：
--
作者：
Perkins,KK;Furneaux,HM;Hurwitz,J
已从HeLa细胞核提取物中分离出三个组分(命名为Ia、Ib和II)，这三个组分是在体外剪接腺病毒和人β-珠蛋白RNA转录本所需的。其中两个组分(Ib和II)在ATP存在下孵育，导致5‘剪接位点前体mRNA的切割和内含子-外显子套索的形成。将组分Ia加入到Ib和Ii的组合中，形成剪接的RNA和内含子套索。当组分II与前体RNA在三磷酸腺苷存在下孵育，所得产物通过蔗糖梯度沉淀，检测到含有前体RNA的30S复合体。组分Ib和组分II的组合导致了55S复合体的产生，该复合体包含作为一个重要的RNA物种的5‘外显子。组分I(包括Ia和Ib)和组分II的结合形成了55S复合体，并在40 S和20 S之间发生物质沉积，其中主要的核糖核酸物种是剪接核糖核酸。
Three fractions (designated Ia, Ib, and II) have been isolated from HeLa cell nuclear extracts that are required for splicing of adenovirus and human beta-globin RNA transcripts in vitro. The incubation of two of the fractions (Ib and II) in the presence of ATP resulted in cleavage of precursor mRNA at the 5' splice site and formation of the intron-exon lariat. Addition of fraction Ia to the combination of Ib and II resulted in the formation of spliced RNA and the intron lariat. When fraction II was incubated with precursor RNA in the presence of ATP and the resulting products were sedimented through sucrose gradients, a 30S complex was detected that contained precursor RNA. The combination of fractions Ib and II resulted in the production of a 55S complex that contained the 5' exon as a prominent RNA species. The combination of fractions I (containing Ia and Ib) and II resulted in the formation of the 55S complex and material sedimenting between 40 S and 20 S, in which the predominant RNA species was spliced RNA.