Zarkower D, Wickens M
Department of Biochemistry, College of Agriculture and Life Sciences, University of Wisconsin-Madison 53706.
EMBO J. 1987 Dec 20;6(13):4185-92. doi: 10.1002/j.1460-2075.1987.tb02765.x.
Complexes form between processing factors present in a crude nuclear extract from HeLa cells and a simian virus 40 (SV40) late pre-mRNA which spans the polyadenylation [poly(A)] site. A specific 'pre-cleavage complex' forms on the pre-mRNA before cleavage. Formation of this complex requires the highly conserved sequence AAUAAA: it is prevented by mutations in AAUAAA, and by annealing DNA oligonucleotides to that sequence. After cleavage, the 5' half-molecule is found in a distinct 'post-cleavage complex'. In contrast, the 3' half-molecule is released. After cleavage and polyadenylation, polyadenylated RNA also is released. De novo formation of the post-cleavage complex requires AAUAAA and a nearby 3' terminus. Competition experiments suggest that a component which recognizes AAUAAA is required for formation of both pre- and post-cleavage complexes.
在来自HeLa细胞的粗核提取物中存在的加工因子与跨越聚腺苷酸化[poly(A)]位点的猿猴病毒40(SV40)晚期前体mRNA之间形成复合物。在切割之前,前体mRNA上会形成一种特定的“切割前复合物”。这种复合物的形成需要高度保守的序列AAUAAA:AAUAAA中的突变以及将DNA寡核苷酸退火到该序列会阻止其形成。切割后,5'半分子存在于一个独特的“切割后复合物”中。相比之下,3'半分子会被释放。切割和聚腺苷酸化后,多聚腺苷酸化的RNA也会被释放。切割后复合物的从头形成需要AAUAAA和附近的3'末端。竞争实验表明,识别AAUAAA的一个组分对于切割前和切割后复合物的形成都是必需的。