RNA的转录后加工(1)分子生物学.pptxVIP

1977年Robert J and Sharp P A分别发现断裂基因（interrupted gene），1993年获得诺贝尔生理学和医学奖 当用RNA与其转录的模板DNA分子杂交时，RNA链取代DNA双链中对应的链，形成R-突环(R-loop);3.7.1 RNA中的内含子;3.7.2 tRNA前体的加工;真核生物tRNA基因的数目比原核生物tRNA基因的数目要大得多。 真核生物的tRNA也成簇排列，并且被间隔区所分开 真核生物tRNA前体的3‘端不含CCA序列，成熟tRNA 3’端的CCA是后加上去的，由核苷酸转移酶催化此反应。 tRNA的修饰成分由特异的修饰酶所催化。真核生物的tRNA除含有修饰碱基外，还有2‘-O-甲基核糖，其含量约为核苷酸的百分之一。 有些tRNA还具有居间序列，需要进一步拼接;3.7.3 原核生物rRNA的加工;真核生物rRNA前体的加工;RNA的拼接共有4种方式： 核mRNA的拼接体的拼接(nuclear mRNA spliceosomal splicing) 类型I自我拼接(group I self-splicing) 类型II自我拼接(group II self-splicing) 核tRNA的酶促拼接（nuclear tRNA enzymatic splicing）;细胞内已发现的RNA剪接有3种（不包括tRNA的加工）。;hnRNA的拼接;此规则不适合于线粒体和叶绿体基因的内含子，也不适合于tRNA和rRNA的内含子;剪接的普遍性;剪接的特异性; pre-mRNA剪接的机制和套索结构;腺苷酸原来已有3’,5’-磷酸二酯键依然存在，加上此2’,5’-磷酸二酯键连接后，在腺苷酸处出现了一个套索。;RNA 剪接由剪接体（splicesome）执行;The snRNPs 在剪接中有3个功能： 识别5 剪接位点和分支点; 把这2个位点集结到一起; 催化或协助催化剪接和连接反应。;U2AF (U2 auxiliary factor), recognizes the polypyrimidine (Py) tract/3 splice site,;snRNA pairing is important in splicing;U1 snRNP initiates splicing;U2 pairs with the branch site;U6 pairs with the 5’-splicing site; 西北农林科技大学 郭泽坤;U4的释放，促进U6和U2配对;U5 snRNP helps to bring the two exons together;U2, U5, U6 bring the reaction group close ;一个矛盾（paradox）;外显子遗漏;The first guard mechanism;The second guard mechanism;how does alternative splicing occur so often? The basic answer is that some splice sites are used only some of the time, leading to the production of different versions of the RNA from different transcripts of the same gene. Alternative splicing can be either constitutive or regulated. In the former case, more than one product is always made from the transcribed gene. In the case of regulated splicing, different forms are generated at different times, under different conditions, or in different cell or tissue types.;正常情况;肌钙蛋白T;两种抗原的比例因剪接相关蛋白SF2/ASF的表达水平而不同。SF2/ASF is an SR protein, when abundant, this protein directs the machinery to favor use of the closest 5 splice site.;Alternative Splicing Is Regulated by Activators and Repressors;Most of activa