分子生物学(中英对照有备注)5.pptVIP

* These experiments showed the presence of introns, but left some questions. Were the introns even transcribed? Or were they transcribed and then removed? In order to verify the existence of RNA splicing, the pre-mRNA for a gene was isolated and hybridized to DNA. Unlike the mRNA, pre-mRNA hybridized perfectly to the DNA without leaving any loops. This meant that introns were present in the pre-mRNA, and that introns are indeed transcribed and then removed later. 这些实验显示了内含子确实存在，但也留下了其它问题。内含子是否也被转录了呢？或者，它们是否先被转录出来然后才被去除呢？为了证实RNA剪接的存在，需要将基因的前体mRNA分离出来与DNA杂交。与mRNA不同，前体mRNA与DNA能完全杂交而不留下任何环。这意味着在前提mRNA中是存在内含子的，因此内含子确实是先被转录出来之后才被去除的。 * But how does a cell know where to cut away this short piece of pre-mRNA? It has been found that there is a polyadenylation signal on the pre-mRNA. This signal has a typical sequence of AAUAAA. This sequence gives an instruction as “to cut the mRNA about 20 nucleotides downstream, near a GU-rich sequence”. 但是，细胞是怎么知道在哪儿切去那一小段的呢？已经发现，在前体mRNA上有聚腺苷酸化信号。该信号的特征性序列是AAUAAA。该序列提示：应在下游约20个核苷酸处靠近富含GU的地方切断。 * If a signal is there to tell where to cut the RNA, then who is there to respond to this signal? Actually, the polyadenylation signal is detected by a cleavage complex. The cleavage complex is composed of four proteins. They are CPSF (cleavage and polyadenylation specificity factor), cleavage stimulation factor protein (CstF) , cleavage factors I and II (CF I and CF II). 如果有了信号告诉说应该在哪儿切断RNA，那么谁来响应这一信号呢？实际上，聚腺苷酸化信号是由切割复合体探测到的。切割复合体由四种蛋白质组成，即CPSF（切割与聚腺苷酸化特异因子）、CstF（切割激活因子）、CF I和CF II（切割因子I和II）。 * Now, we are ready to look at the process of polyadenylation. First, after the RNA polymerase has transcribed the polyadenylation signal AAUAAA, CPSF will bind to the AAUAAA sequence. At the same time, CstF binds to the G-U rich sequence downstream. Then, CF I and CF II bind and cut the mRNA between the AAUAAA sequence and the G-U rich sequence to produce a terminus for polyadenylation. Finally, a specialized RNA polymerase called poly(A) polym