14. RNA的生物合成与加工-20131205.ppt

四膜虫rRNA 四膜虫rRNA内含子的二级结构 四膜虫rRNA的剪接采用自我剪接方式 5′-端核苷酸序列 核酶 核酶：RNA本身具有催化活性，此种由RNA发挥催化作用的酶，称为核酶。 最简单的核酶呈槌头结构。 最简单的核酶二级结构——槌头状结构 (hammerhead structure) 底物部分 通常为60个核苷酸左右 同一分子上包括有催化部份和底物部份 催化部份和底物部份组成锤头结构 除rRNA外，tRNA、mRNA的加工也可采用自我剪接方式。 核酶研究的意义 核酶的发现，对中心法则作了重要补充； 核酶的发现是对传统酶学的挑战； 对进化的研究有帮助； 利用核酶的结构设计合成人工核酶,应用于疾病的治疗。 G OH 3′ G 5′ OH 5′ 3′ G OH 414 G 399 5′ 3′ 395 5′ 3′ E1 E2 I 四膜虫RNA的自我剪接 5′ 3′ L19RNA 具有催化活性的片段 附录 首先发现的核酶：L19 RNA 具催化活性： 核苷酸转移酶 2CpCpCpCpCp ? Cp6 + Cp4 磷酸二酯酶 CpCpCpCpC ? Cp4+ pC 磷酸转移酶 Cp4Cp ——? Cp4C+UpCpUp 限制性内切酶 RNA-CUCU ? RNA’+CUCU 反应具有专一性：只催化RNA，对DNA? 有竞争性抑制剂：dpC5，Ki=260?mol/L UpCpU 转录与复制的相似之处 都以DNA为模板 需要核苷酸作原料，从5’到3’延长；生成磷酸二酯键以连接核苷酸 都遵从碱基配对规律 都需要依赖DNA的聚合酶 产物都是很长的多核苷酸 转录和复制的区别 * FIGURE 26-32 Extension of the central dogma to include RNA-dependent synthesis of RNA and DNA. * FIGURE 26-27 Processing of tRNAs in bacteria and eukaryotes. The yeast tRNATyr (the tRNA specific for tyrosine binding; see Chapter 27) is used to illustrate the important steps. The nucleotide sequences shown in yellow are removed from the primary transcript. The ends are processed first, the 5′ end before the 3′ end. CCA is then added to the 3′ end, a necessary step in processing eukaryotic tRNAs and those bacterial tRNAs that lack this sequence in the primary transcript. While the ends are being processed, specific bases in the rest of the transcript are modified (see Figure 26-23). For the eukaryotic tRNA shown here, the final step is splicing of the 14 nucleotide intron. Introns are found in some eukaryotic tRNAs but not in bacterial tRNAs. * FIGURE 26-30 Secondary structure of the self-splicing rRNA intron of Tetrahymena. Intron sequences are shaded yellow, exon sequences green. Each thick yellow line represents a bond between neighboring nucleotides in a continuous sequence (a device necessitated by showing this complex molecule in two dimensions; similarly, an oversize blue line between a C and G residue indicates normal base pairing); all nucleotides are shown. T