Evidence that a nuclear matrix protein participates in premessenger RNA splicing.
Evidence that a nuclear matrix protein participates in premessenger RNA splicing.
复制标题
核基质蛋白参与前信使 RNA 剪接的证据。
DOI：
10.1016/0014-4827(89)90255-3
复制
发表时间：
1989
影响因子：
3.7
通讯作者：
Berget,SM
中科院分区：
文献类型：
--
作者：
Smith,HC;Harris,SG;Zillmann,M;Berget,SM
用抗分离大鼠肝细胞核基质的抗体，并与65 kDa HeLa细胞核基质蛋白(IGA-65)交叉反应，研究了核基质蛋白在前信使RNA剪接中的作用。IGA-65是一种核内基质组分，在提取液制备过程中，通过内源性核糖核酸酶、EDTA和DTT的作用，可以作为核剪接体提取液的一种成分进行溶解。剪接提取物与抗IGA-65抗体(抗IGA-65)预育抑制外源腺病毒前体RNA的体外剪接。此外，抗IGA-65抗体可抑制前体RNA组装成活性剪接体复合体，提示IGA-65在早期剪接体组装过程中起作用。在蛋白质凝胶上，剪接提取物中存在的IGA-65与已知的U-SnRNP和hnRNP蛋白是不同的。此外，剪接提取物在天然凝胶上的电泳表明，IGA-65存在于不同于U-SnRNPs或hnRNP C蛋白的蛋白质复合体中。这些数据支持将含有IGA-65的复合体确定为参与前mRNA剪接的核因子，并由此表明核基质在体内处理过程中的作用。
The role of nuclear matrix proteins in premessenger RNA splicing has been investigated using antibodies raised against isolated rat liver nuclear matrix and cross-reactive with a 65-kDa HeLa cell nuclear matrix protein (IGA-65). IGA-65 is an internal nuclear matrix component which can be solubilized as a component of nuclear splicing extracts, by the action of endogenous ribonucleases, EDTA, and DTT during extract preparation. Preincubation of splicing extract with antibodies against IGA-65 (anti-IGA-65) inhibitedin vitrosplicing of exogenous adenovirus precursor RNA. Furthermore, assembly of precursor RNA into active spliceosome complexes was inhibited by pretreatment of extracts with anti-IGA-65, suggesting a role for IGA-65 during early spliceosome assembly. The IGA-65 present in splicing extracts was distinguishable from known U-snRNP and hnRNP proteins on protein gels. Furthermore, electrophoresis of splicing extract on native gels indicated that IGA-65 was present in protein complexes different from those containing U-snRNPs or hnRNP C protein. The data support identification of complexes containing IGA-65 as nuclear factors involved in pre-mRNA splicing and, by extension, suggest a role for the nuclear matrix during processingin vivo.