重组RNA聚合酶II复合体,以便在体内复制感染性非编码RNA
bioRxiv : the preprint server for biology
|June 4, 2025
概括
这项研究揭示了必不可少的RNA聚合酶II (Pol II) 酶如何在体内重组成一个较小的7个子单元复合体,以转录病毒性RNA. 转录因子IIIA的一个特定变体促进了这种新型RNA-依赖RNA聚合酶活性.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 结构生物学 结构生物学
背景情况:
- 依赖DNA的RNA聚合酶 (DdRPs) 具有依赖RNA的RNA聚合酶 (RdRP) 活性,被病毒病毒和型肝炎病毒利用.
- 这种RdRP活性背后的分子机制在很大程度上是未知的.
- 病毒RNA复制依赖于宿主RNA聚合酶II (Pol II) 和其RdRP功能.
研究的目的:
- 在Pol II中阐明RNA-依赖RNA聚合酶活性的分子基础.
- 在 viroid RNA 合成过程中研究 Pol II 转录复合物的结构组织.
- 为了确定参与Pol II介导的病毒转录的宿主因素.
主要方法:
- 在植物分析中,Pol II亚单元参与了土豆管病毒 (PSTVd) 转录.
- 使用AlphaFold3进行结构建模,以预测重建的Pol II,PSTVd RNA和转录因子IIIA的复合体.
- 对PSTVd循环8的突变分析,以评估其在TFIIIA-PSTVd相互作用中的作用.
主要成果:
- 在 PSTVd 转录中,Pol II 在体内从12个子单元重组为7个子单元复合体.
- 特定的子单元 (Rpb4,Rpb5,Rpb6,Rpb7,Rpb9) 不是PSTVd转录所必需的.
- 转录因子IIIA (TFIIIA-7ZF) 的一个拼接变体与PSTVdRNA和重塑的Pol II (Rpb2) 相互作用.
- AlphaFold3建模预测了TFIIIA-7ZF与PSTVd的N端和Rpb2的结合,并在PSTVd循环8上确定了一个关键的曲区域.
- 破坏PSTVd循环8的灵活性会损害TFIIIA-7ZF的相互作用.
结论:
- 病毒转录在体内涉及一种异质的,重塑的基本Pol II酶的形式.
- TFIIIA-7ZF作为一个关键的适配器,桥接PSTVdRNA和重建的PolII.
- 结构洞察力揭示了特定RNA结构元素 (循环8) 对酶-RNA相互作用和转录启动的重要性.
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