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首页> 外文期刊>Molecular and Cellular Biology >Sequences downstream of AAUAAA signals affect pre-mRNA cleavage and polyadenylation in vitro both directly and indirectly.
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Sequences downstream of AAUAAA signals affect pre-mRNA cleavage and polyadenylation in vitro both directly and indirectly.

机译：AAUAAA信号下游的序列直接或间接地在体外影响前mRNA的切割和聚腺苷酸化。

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摘要

To investigate the role of sequences lying downstream of the conserved AAUAAA hexanucleotide in pre-mRNA cleavage and polyadenylation, deletions or substitutions were constructed in polyadenylation signals from simian virus 40 and adenovirus, and their effects were assayed in both crude and fractionated HeLa cell nuclear extracts. As expected, these sequences influenced the efficiency of both cleavage and polyadenylation as well as the accuracy of the cleavage reaction. Sequences near or upstream of the actual site of poly(A) addition appeared to specify a unique cleavage site, since their deletion resulted, in some cases, in heterogeneous cleavage. Furthermore, the sequences that allowed the simian virus 40 late pre-RNA to be cleaved preferentially by partially purified cleavage activity were also those at the cleavage site itself. Interestingly, sequences downstream of the cleavage site interacted with factors not directly involved in catalyzing cleavage and polyadenylation, since the effects of deletions were substantially diminished when partially purified components were used in assays. In addition, these sequences contained elements that could affect 3'-end formation both positively and negatively.
机译：为了研究保守的AAUAAA六核苷酸下游序列在前mRNA切割和聚腺苷酸化中的作用，构建了猿猴病毒40和腺病毒的聚腺苷酸化信号的缺失或替代，并在粗制和分级HeLa细胞核提取物中检测了它们的作用。如所预期的，这些序列影响切割和聚腺苷酸化的效率以及切割反应的准确性。在poly（A）添加的实际位点附近或上游的序列似乎指定了一个独特的切割位点，因为在某些情况下，其缺失会导致异源切割。此外，允许猿猴病毒40晚期pre-RNA通过部分纯化的切割活性优先被切割的序列也是在切割位点本身的那些序列。有趣的是，切割位点下游的序列与不直接参与催化切割和聚腺苷酸化的因子相互作用，因为当在测定中使用部分纯化的组分时，缺失的影响大大降低。另外，这些序列包含可能正向和负向影响3'末端形成的元件。

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