在nsp12-nsp8接口的特定部位的点突变极大地影响SARS-CoV-2的RNA聚合活性
Cristina Ferrer-Orta1, Sergi Vázquez-Monteagudo1, Diego S Ferrero1
1Structural and Molecular Biology Department, Institut de Biologia Molecular de Barcelona, Consejo Superior de Investigaciones Científicas, Barcelona 08028, Spain.
概括
在SARS-CoV-2的突变.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 严重急性呼吸系统综合征 冠状病毒2 (SARS-CoV-2) 依赖RNA的RNA聚合酶 (RdRp) 对病毒复制至关重要.
- 了解突变对RdRp活性的影响,是开发抗病毒策略的关键.
研究的目的:
- 研究SARS-CoV-2 nsp12中特定氨基酸替代物对RdRp活性的功能影响.
- 确定nsp12-nsp8接口附近的突变如何影响RNA聚合.
主要方法:
- 位点定向突变发生,在nsp12中引入特定的氨基酸替代物.
- 试验室初始延长试验测量RdRp活性.
- 对影响nsp12-nsp8疏水群的突变进行分析.
主要成果:
- 在聚合酶中心腔 (D499G,M668V,V560A) 中的替代物没有显著改变RdRp活性.
- 在nsp12-nsp8接口附近的突变 (P323L,L372F,L372P,V373A,L527H) 显著影响了RNA聚合.
- L372F略增强了活性,而L372P,L527H和V373A通过削弱疏水相互作用来减少了活性.
结论:
- nsp12-nsp8接触接口在调节SARS-CoV-2RNA合成方面发挥着至关重要的作用.
- 影响这种接口的特定突变可以显著损害病毒RdRp活动,提供潜在的抗病毒标.
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