Cotten M, Schaffner G, Birnstiel M L
Institute for Molecular Pathology, Vienna, Austria.
Mol Cell Biol. 1989 Oct;9(10):4479-87. doi: 10.1128/mcb.9.10.4479-4487.1989.
A comparative analysis of ribozyme, antisense RNA, and antisense DNA inhibitors of the in vitro small nuclear ribonucleoprotein U7-dependent histone pre-mRNA processing reaction was performed. RNA molecules complementary to the U7 sequence inhibited in vitro processing of histone pre-mRNA at a sixfold excess over U7. Single-stranded DNA complementary to the entire U7 sequence inhibited the reaction at a 60-fold excess over U7, while a short, 18-nucleotide DNA molecule complementary to the 5' end of U7 inhibited the processing reaction at a 600-fold excess. A targeted ribozyme was capable of specifically cleaving the U7 small nuclear ribonucleoprotein in a nuclear extract and inhibited the U7-dependent processing reaction, but in our in vitro system it required a 1,000-fold excess over U7 for complete inhibition of processing.
对核酶、反义RNA和反义DNA抑制剂在体外小核核糖核蛋白U7依赖性组蛋白前体mRNA加工反应中的作用进行了比较分析。与U7序列互补的RNA分子在比U7过量六倍时抑制组蛋白前体mRNA的体外加工。与整个U7序列互补的单链DNA在比U7过量60倍时抑制该反应,而与U7 5'端互补的短的18个核苷酸的DNA分子在比U7过量600倍时抑制加工反应。一种靶向核酶能够在核提取物中特异性切割U7小核核糖核蛋白并抑制U7依赖性加工反应,但在我们的体外系统中,它需要比U7过量1000倍才能完全抑制加工。