分子生物学第五章分析.ppt

Now, how does capping take place? Actually, three enzymes are involved in the capping process. First, the RNA triphosphatase removes one phosphate from the 5’ end of the pre-mRNA. Second, guanylyl transferase adds a guanine nucleotide to make 5’-5’ linkage. Finally, methyl transferase adds a methyl group to the guanine nucleotide. 那么，加帽是如何进行的呢？实际上，加帽过程需要有三种酶参与。首先，RNA三磷酸酶首先从前体mRNA的5’端移去一个磷酸；之后鸟苷酸转移酶加上一个鸟嘌呤核苷酸以形成5’-5’键；最后，甲基转移酶在鸟嘌呤核苷酸上加上一个甲基。 These three enzymes stay together around the C-terminal domain of the Rpb1 subunit of RNA polymerase II. They act together to complete the capping process. Because this C-terminal domain is very important, it is often called ‘the CTD.’ 这三种酶在RNA聚合酶II开始转录前体mRNA的时候聚集在它的Rpb1亚基的C末端功能域中。由于这一功能域的重要性，它常常就被称为‘该CTD’。 Somebody may ask why a cap is added to mRNA. Actually, the cap structure is very useful to mRNA. It is known to have four functions. First, it helps prevent degradation. Second, it helps transport the RNA into cytoplasm. Thirdly, it enhances translation. And finally, it helps remove the first intron. 有人可能要问为什么要在mRNA上加帽，实际上，帽结构对mRNa是非常有用的。帽结构具有四种主要功能。 第一，它防止RNA被降解；第二，它帮助RNA转运到细胞质中；第三，它能增强转译；第四，它帮助去除第一个内含子。 First, it helps to prevent degradation of the mRNA at the 5’ end. RNAs in the cell can be rapidly degraded by enzymes called ribonucleases. However, these enzymes are generally not capable of degrading the triphosphate bond introduced by the cap structure. 首先，它对防止mRNA在5’端发生降解有帮助。细胞中的RNA会被称为核糖核酸酶的酶迅速降解掉。然而，这些酶一般不能降解帽结构中的三磷酸键。 真核生物mRNA5′端有m7G5ppp5′Nm2′pNp…的特殊结构，它抗5′～核酸外切酶的降解，与蛋白质合成的正确起始作用有关。这一特殊结构称为帽子结构。（ Second, the cap structure helps the RNA transcript to pass through the selective pores of the nuclear membrane and into the cytoplasm. This is important because transcription occurs in the nucleus, but the ribosomes that translate mRNA are located in the cytoplasm. 第二，帽结构帮助RNA转录产物穿过核膜的选择性孔道而进入细胞质。这一点很重要，因为转录发生在细胞核，而转译mRNA的核糖体位于细胞质中。 Thirdly, the cap structure enhances translation. In order to bind the ribosome