通过SARS-CoV-2聚合酶和S759A耐药性的选择性remdesivir整合的结构基础
Kalyan Das1, Calvin Gordon2, Mizar Oliva1
1KU Leuven.
Research square
|November 24, 2025
概括
雷梅西维尔三酸盐 (RTP) 被SARS-CoV-2RNA依赖RNA聚合酶 (RdRp) 纳入,但连续的纳入受到ATP存在和结构刚性的限制. 这一发现影响了新型抗病毒核酸类类似物的设计.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 药物发现 药物发现 药物发现
背景情况:
- 核糖类类似物是有效的抗病毒药物.
- 雷姆德西维尔 (一种核类型) 通过延迟链终结或模板链结合来抑制RNA合成.
研究的目的:
- 阐明通过SARS-CoV-2RNA依赖RNA聚合酶 (RdRp) 结合remdesivir三酸盐 (RTP) 的机制.
- 了解限制RTP结合的因素,并为新抗病毒核酸类同类的设计提供信息.
主要方法:
- 酶性检测试验 酶性检测试验
- 质谱测量质量谱测量
- 低温电子显微镜 (cryo-EM) 用于确定RdRp结构.
- 位点定向的突变发生 (S759A突变)
主要成果:
- 在SARS-CoV-2中,RdRp优先结合RTP而不是ATP,即使ATP超过10倍.
- 当ATP存在时,连续的RTP整合是不利的,因为弹性remdesivir:UMP基对阻碍了转位.
- 雷梅西维尔的占用率仅限于复制RNA链中可用位置的<16%.
- S759A RdRp突变体表现出对RTP的抗性,这与原始3'-end核酸重新定位和改变的核糖环形状有关.
结论:
- 雷梅西维尔由SARS-CoV-2 RdRp的结合受到核酸竞争和聚合酶结构动态的调节.
- 这些发现提供了关于雷梅西维尔的作用机制的见解,并指导了下一代核类类似物的开发,并改进了抑制策略.
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