用生物素化的2'-O-甲基RNA寡核苷酸对人U4/U6核小核糖核蛋白进行反义探测。

Antisense probing of the human U4/U6 snRNP with biotinylated 2'-OMe RNA oligonucleotides.

作者信息

Blencowe B J, Sproat B S, Ryder U, Barabino S, Lamond A I

机构信息

European Molecular Biology Laboratory, Heidelberg Federal Republic of Germany.

出版信息

Cell. 1989 Nov 3;59(3):531-9. doi: 10.1016/0092-8674(89)90036-6.

DOI:10.1016/0092-8674(89)90036-6

PMID:2478298

Abstract

We have used antisense 2'-OMe RNA oligonucleotides carrying four 5'-terminal biotin residues to probe the structure and function of the human U4/U6 snRNP. Nine oligonucleotides, complementary to multiple regions of U4 and U6 snRNAs, bound stably and specifically to U4/U6 snRNP. This allowed for efficient and selective removal of U4/U6 from HeLa cell nuclear extracts. Binding of oligonucleotides to certain snRNA domains inhibited splicing and affected the U4-U6 interaction. Pre-mRNA and splicing products could also be affinity-selected through binding of the oligonucleotides to U4/U6 snRNPs in splicing complexes. The results suggest that U4 snRNP is not released during spliceosome assembly.

摘要

我们使用了携带四个5'-末端生物素残基的反义2'-OMe RNA寡核苷酸来探测人U4/U6核小核糖核蛋白颗粒（snRNP）的结构和功能。九条与U4和U6小核RNA（snRNA）多个区域互补的寡核苷酸稳定且特异地结合到U4/U6 snRNP上。这使得从HeLa细胞核提取物中高效且选择性地去除U4/U6成为可能。寡核苷酸与某些snRNA结构域的结合抑制了剪接并影响了U4-U6相互作用。前体mRNA和剪接产物也可以通过寡核苷酸与剪接复合物中U4/U6 snRNPs的结合进行亲和选择。结果表明U4 snRNP在剪接体组装过程中不会释放。