Grabowski P J, Seiler S R, Sharp P A
Cell. 1985 Aug;42(1):345-53. doi: 10.1016/s0092-8674(85)80130-6.
A multicomponent complex termed spliceosome (splicing body) is unique to the splicing of messenger RNA precursors in vitro. This 60S RNA-protein complex contains RNAs from the previously characterized bipartite splicing intermediate, the 5' exon RNA, and the lariat intervening sequence-3' exon RNA, as well as some intact 455 nucleotide precursor RNA. This complex contains snRNPs, particularly U1 RNP, as shown by immunoprecipitation with specific antisera. Formation of the 60S complex appears to be an early and essential step in splicing, because the 60S complex forms during the early stage, or lag time, of the reaction before the first covalent modification, cleavage at the 5' splice site of precursor RNA. The 60S complex forms only under conditions that permit splicing; both ATP and a precursor RNA containing authentic 5' and 3' splice sites are required for formation, while antiserum specific for U1 RNP inhibits its formation. RNA within the 60S complex, predominantly precursor RNA, was chased into products with accelerated kinetics and more complete conversion than purified precursor RNA.
一种称为剪接体(拼接体)的多组分复合物在体外对信使核糖核酸前体的剪接具有独特性。这种60S核糖核蛋白复合物包含来自先前已鉴定的二分体剪接中间体的RNA、5'外显子RNA和套索状内含序列-3'外显子RNA,以及一些完整的455个核苷酸的前体RNA。如用特异性抗血清进行免疫沉淀所示,该复合物含有小核核糖核蛋白颗粒,特别是U1核糖核蛋白颗粒。60S复合物的形成似乎是剪接过程中一个早期且必不可少的步骤,因为60S复合物在反应的早期阶段或延迟期形成,即在第一个共价修饰、前体RNA的5'剪接位点切割之前。60S复合物仅在允许剪接的条件下形成;形成过程需要ATP和含有真实5'和3'剪接位点的前体RNA,而针对U1核糖核蛋白颗粒的抗血清会抑制其形成。60S复合物中的RNA,主要是前体RNA,与纯化的前体RNA相比,以更快的动力学被追踪到产物中,并且转化更完全。