分子生物学Chapter-5.pptVIP

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。 聚腺苷酸化并不是在转录产物前体mRNA本身的尾部发生的。实际上，要将前体mRNA上先切去一小段，之后poly(A)尾才被加上去 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) polymerase comes and adds a lot of As at this site. After cleavage of the transcript, CPSF remains bound at the AAUAAA site. A specialized RNA polymerase called poly(A) polymerase is then required to add adenosine nucleotides. Poly(A) polymerase begins slowly at first, and is eventually accompanied by a protein called poly(A)-binding protein (PBP). PBP can help poly(A) polymerase to work more efficiently. 现在我们可以开始看聚腺苷酸化过程了。首先，在RNA聚合酶转录出了聚腺苷酸化信号AAUAAA后，CPSF将结合到AAUAAA序列上，与此同时CstF会结合到富含GU的下游序列上。之后，CF I和CF II在AAUAAA序列和富含GU的序列之间将mRNA切断，从而为聚腺苷酸化产生出所需要的末端。最后，一种称为poly(A)聚合酶的特殊RNA聚合酶将腺嘌呤