6++RNA+的剪接分析.pptVIP

RNA Splicing;Figure 13-1; 多数真核基因都是断裂的，其编码序列由非编码序列隔开 Exons (外显子): the coding sequences Introns (内含子) : the intervening sequences RNA splicing: the process by which introns are removed from the pre-mRNA. Alternative splicing (可变剪接): some pre-mRNAs can be spliced in more than one way , generating alternative mRNAs. 60% of the human genes are spliced in this manner.;Topic 1 : RNA剪接的化学反应;RNA的序列决定剪接位点;Figure 13-2;5’splice site (5’剪接位点): the exon-intron boundary at the 5’ end of the intron 3’ splice site (3’剪接位点): the exon-intron boundary at the 3’ end of the intron Branch point site (分枝位点): an A close to the 3’ end of the intron, which is followed by a polypyrimidine tract (Py tract).;两相邻的外显子连接起来，内含子以套马索(Lariat)的结构被除去;Figure 13-3;丁字路口结构 Three-way junction;Step 2: The OH of the 5’ exon attacks the phosphoryl group at the 3’ splice site. As a consequence, the 5’ and 3’ exons are joined and the intron is liberated in the shape of a lariat.;Figure 13-3;来至不同RNA分子的外显子可以通过反式剪接(Trans-splicing)连接起来;;Topic 2：剪接机器 ;RNA剪接由一个大的叫剪接体(spliceosome)的复合体执行;5 RNAs (U1, U2, U4, U5, and U6, 100-300 nt) 叫核内小RNA(small nuclear RNAs, snRNAs). snRNA和蛋白的复合体叫 核内小分子核糖核蛋白 small nuclear ribonucleoproteins (snRNP, 发音 “snurps”). 剪接体是最大的snRNP，在剪接的不同阶段它的组成会有变化 ;1. 识别 the 5’ splice site and the branch site. 2. 将这两个位点带到一起. 3. 催化 (or 帮助催化) RNA 断裂. RNA-RNA, RNA-protein and protein-protein interactions are all important during splicing.;Figure 13-6;Topic 3：剪接途径;剪接体的装配, 重排, 和催化 : the splicing pathway (Fig. 13-8);装配 step 2 1. U2 binds to the branch site, and then A complex is formed. 2. The base-pairing between the U2 and the branch site is such that the branch site A is extruded(Figure 13-6). This A residue is available to react with the 5’ splice site.;Figure 13-8;装配 step 3 1. U4, U5 and U6 form the tri-snRNP Particle. 2. With the entry of the tri-snRNP, the A complex is converted into the B complex.;Figure 13-8;装配 step 4 U1 leaves the complex, and U6 replaces it at the 5’ splice site. U4 is relea