Parker R, Siliciano P G, Guthrie C
Cell. 1987 Apr 24;49(2):229-39. doi: 10.1016/0092-8674(87)90564-2.
The U2 snRNP binds to the site of branch formation during splicing of mammalian pre-mRNA in vitro. In Saccharomyces cerevisiae the branch site is within the so-called TACTAAC box (UACUAAC box), an absolutely conserved intron sequence required for splicing. Based on the identification and sequence of a U2 analogue in yeast, a specific base pairing interaction between the UACUAAC box and a highly conserved region of this snRNA can be proposed. To test this hypothesis, we have taken advantage of two mutations constructed previously in the UACUAAC box of an actin-HIS4 fusion. These mutant strains were transformed with stable plasmids bearing U2-like snRNAs into which changes predicted to restore base pairing had been introduced. Allele-specific suppression of biological and biochemical phenotypes was observed in both cases. Recognition of the UACUAAC box thus relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue.
在体外剪接哺乳动物前体mRNA的过程中,U2小核核糖核蛋白颗粒(snRNP)结合到分支形成位点。在酿酒酵母中,分支位点位于所谓的TACTAAC框(UACUAAC框)内,这是剪接所需的一个绝对保守的内含子序列。基于在酵母中对U2类似物的鉴定和序列分析,可以推测UACUAAC框与该小核RNA(snRNA)的一个高度保守区域之间存在特定的碱基配对相互作用。为了验证这一假设,我们利用了先前在肌动蛋白-HIS4融合基因的UACUAAC框中构建的两个突变。用携带U2样snRNA的稳定质粒转化这些突变菌株,这些质粒中已引入了预测可恢复碱基配对的变化。在这两种情况下均观察到了生物和生化表型的等位基因特异性抑制。因此,对UACUAAC框的识别至少部分依赖于与酵母U2类似物的沃森-克里克碱基配对。