Hamm J, Dathan N A, Scherly D, Mattaj I W
European Molecular Biology Laboratory, Heidelberg, FRG.
EMBO J. 1990 Apr;9(4):1237-44. doi: 10.1002/j.1460-2075.1990.tb08231.x.
Domains of U1 snRNA which are functionally important have been identified using a splicing complementation assay in Xenopus oocytes. Mutations in, and deletions of, all three of the hairpin loop structures near the 5' end of the RNA are strongly deleterious. Similarly, mutation of the Sm binding site abolishes complementation activity. Analysis of the protein binding properties of the mutant U1 snRNAs reveals that three of the functionally important domains, the first two hairpin loops and the Sm binding site, are required for interaction with U1 snRNP proteins. The fourth functionally important domain does not detectably affect snRNP protein binding and is not evolutionarily conserved. All of the deleterious mutations are shown to have similar effects on in vivo splicing complex formation.
利用非洲爪蟾卵母细胞中的剪接互补分析，已确定了U1 snRNA具有功能重要性的结构域。RNA 5'端附近的所有三个发夹环结构发生突变和缺失均具有强烈的有害性。同样，Sm结合位点的突变会消除互补活性。对突变型U1 snRNAs的蛋白质结合特性分析表明，与U1 snRNP蛋白质相互作用需要三个具有功能重要性的结构域，即前两个发夹环和Sm结合位点。第四个具有功能重要性的结构域对snRNP蛋白质结合没有可检测到的影响，并且在进化上不保守。所有有害突变对体内剪接复合体形成均显示出相似的影响。
