流感聚合酶与Pol IICTD之间的基本相互作用的结构基础
Maria Lukarska1, Guillaume Fournier2,3,4, Alexander Pflug1
1European Molecular Biology Laboratory, Grenoble Outstation, 71 Avenue des Martyrs, CS 90181, 38042 Grenoble Cedex 9, France.
Nature
|December 22, 2016
概括
流感聚合酶通过其PA子单元的特定位点与细胞RNA聚合酶II的C终端域 (CTD) 结合. 破坏这种相互作用会损害病毒的转录和复制,这表明它是抗病毒药物的目标.
科学领域:
- 病毒学
- 结构生物学
- 分子生物学
背景情况:
- 流感聚合酶 (FluPol) 对于病毒RNA转录和复制至关重要.
- FluPol 与细胞RNA聚合酶II (Pol II) 相互作用,以获得mRNA转录的5'- capped原始体.
研究的目的:
- 阐明流感聚合酶与Pol II C终端域 (CTD) 相互作用的结构基础.
- 研究这种相互作用对病毒转录和复制的功能意义.
主要方法:
- 蝙蝠流感A聚合酶与Pol IICTD模拟的联合晶体.
- 在PA子单元中确定CTD结合残留物的位点定向突变.
- 在体外RNA合成测试和基于细胞的小基因组测试.
- 复合流感病毒的救援和特征.
主要成果:
- 晶体结构揭示了PA子单元中的两个明显的素-5 (SeP5) 结合位,容纳了多个CTD重复.
- 在实验室中,SeP5结合残留物的突变使CTD结合减弱,但在基于细胞的测定中显著降低了聚合酶活性.
- 在这些部位突变的重组病毒是减弱的和基因不稳定的,其中一些突变与已知的毒性因子相关.
结论:
- 流感聚合酶直接与Pol II CTD结合,特别是在SeP5位点,对于有效的病毒转录至关重要.
- 在流感聚合酶上发现的CTD结合部位是开发新型抗病毒疗法的潜在目标.
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