Hamm J, van Santen V L, Spritz R A, Mattaj I W
European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
Mol Cell Biol. 1988 Nov;8(11):4787-91. doi: 10.1128/mcb.8.11.4787-4791.1988.
The binding of the U1 small nuclear ribonucleoprotein (snRNP)-specific proteins C, A, and 70K to U1 small nuclear RNA (snRNA) was analyzed. Assembly of U1 snRNAs from bean and soybean and a set of mutant Xenopus U1 snRNAs into U1 snRNPs in Xenopus egg extracts was studied. The ability to bind proteins was analyzed by immunoprecipitation with monospecific antibodies and by a protein-sequestering assay. The only sequence essential for binding of the U1-specific proteins was the conserved loop sequence in the 5' hairpin of U1. Further analysis suggested that protein C binds directly to the loop and that the assembly of proteins A and 70K into the RNP requires mainly protein-protein interactions. Protein C apparently recognizes a specific RNA sequence rather than a secondary structural element in the RNA.
分析了U1小核核糖核蛋白(snRNP)特异性蛋白C、A和70K与U1小核RNA(snRNA)的结合情况。研究了来自菜豆和大豆的U1 snRNA以及一组突变的非洲爪蟾U1 snRNA在非洲爪蟾卵提取物中组装成U1 snRNP的过程。通过用单特异性抗体进行免疫沉淀和蛋白质隔离试验分析了结合蛋白的能力。U1特异性蛋白结合所必需的唯一序列是U1 5'发夹结构中的保守环序列。进一步分析表明,蛋白C直接与环结合,而蛋白A和70K组装到核糖核蛋白中主要需要蛋白质-蛋白质相互作用。蛋白C显然识别特定的RNA序列而非RNA中的二级结构元件。