分子生物学课件chapter7 RNA 转录课件.ppt

* ? 转酯反应是磷酸酯键的直接转移 * a、 G与其结合位点以氢键相结合，3’—OH处于向 5’拼接点U的 3’磷酸酯键发动亲核进攻的位置上，5’拼接点断裂 b、 与此类似，5’拼接点断裂后，其 3’—OH也正好处于向 3’拼接点G进攻的有利位置 * U1和U2共同作用使内元的 5’端和 3’端带到一起同时U1与 3’拼接点配对 * 分支点的A的 2’ –OH攻击 5’拼接点G，内元 剪切与套索的形成同步，释放左侧内元 左侧外元的 3’-OH攻击3’拼接点G，内元 剪切（lariat 切 除）与exons连接同步 * * mRNA转录后加帽和加尾（核内）的同时，内元的拼接也在进行，然后转运到胞浆 反式剪接的情况较为稀少，较典型的例子是锥虫表面糖蛋白基因VSG(variable surface glycoprotein)，线虫的肌动蛋白基因（actin genes），和衣藻（chlamydomonas）叶绿体DNA中含有的psa基因。 Trans-splicing 锥虫的许多mRNA都含有这个 35bases 序列,称其为前导序列（leader sequence).前导序列是其基因中所没有的.这种前导序列来源于基因组中位于别处的重复序列（200个）所转录的小片段RNA，每个重复单位长135nt，前面35nt的前导序列可以通过转酯反应加到 mRNA的5′端。 splice leader( SL) Alternative splicing An example of alternative splicing Alternative splicing of SV40 T-antigen The possible mechanisms of alternative splicing 1. Major and minor splicesome 2. nonsense-mediated decay 3. A novel strategy used by Dscam Exon 6 Drosophila Dscam gene- encodes cell surface proteins of the Ig superfamily Alternative splicing is regulated by activators and repressors Regulation of alternative splicing determine the sex of the flies Exon shuffling the significance of broken genes in eukaryotes ? Δ Alternative splicing Δ Reshuffling exons Three evidences 1. The borders between exons and introns within a given gene often coincide with the boundaries between domains within the protein encoded by that gene. 2. Many genes have apparently arisen during evolution in part via exon duplication and divergence. 3. Related exons are sometimes found in otherwise unrelated genes. There are numerous examples of proteins made up of highly related domains used in various combinations, encoded by genes made up of shuffled exons. RNA编辑（RNA editing） RNA editing is a process in which information changes at the level of mRNA. e.g. DNA正链序列： GA G A A mRNA 序列： GAU UGU AUA 蛋白质 序列： Asp Cys Ile In 1986, Rob Benne and his colleagues discovered that the sequence of the cytochrome oxidase(COII)