Mapping inhibitory sites on the RNA polymerase of the 1918 pandemic influenza virus using nanobodies.
Mapping inhibitory sites on the RNA polymerase of the 1918 pandemic influenza virus using nanobodies.
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DOI：
10.1038/s41467-021-27950-w
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发表时间：
2022-01-11
影响因子：
16.6
通讯作者：
Grimes JM
中科院分区：
文献类型：
--
作者：
Keown JR;Zhu Z;Carrique L;Fan H;Walker AP;Serna Martin I;Pardon E;Steyaert J;Fodor E;Grimes JM
甲型流感病毒引起季节性流行病和全球性大流行病，给医疗保健系统带来相当大的负担。流感病毒复制周期的中心是病毒RNA依赖性RNA聚合酶，其转录和复制病毒RNA基因组。聚合酶经历构象重排并与病毒和宿主蛋白相互作用以执行这些功能。在这里，我们确定了1918年流感病毒聚合酶的转录酶和复制酶构象的结构，使用冷冻电子显微镜（冷冻EM）。然后，我们在结构和功能上验证了单域纳米抗体与1918年大流行性流感病毒聚合酶的结合。结合这些功能和结构数据，我们确定了五个网站的聚合酶是敏感的抑制纳米抗体。我们建议，在这些网站的纳米抗体的结合，防止聚合酶从假设特定的功能构象或与病毒或宿主因子的相互作用。该聚合酶在甲型流感亚型中高度保守，表明这些位点是潜在流感抗病毒药物开发的有效靶点。流感病毒携带其自身的高度保守的RNA依赖性RNA聚合酶，并且是有希望的抗病毒靶标。结合功能和结构数据，Keown等人验证了纳米抗体对1918年大流行性H1N1流感病毒株聚合酶复合物的抑制作用，并鉴定了体外干扰聚合酶活性的敏感位点。
Influenza A viruses cause seasonal epidemics and global pandemics, representing a considerable burden to healthcare systems. Central to the replication cycle of influenza viruses is the viral RNA-dependent RNA polymerase which transcribes and replicates the viral RNA genome. The polymerase undergoes conformational rearrangements and interacts with viral and host proteins to perform these functions. Here we determine the structure of the 1918 influenza virus polymerase in transcriptase and replicase conformations using cryo-electron microscopy (cryo-EM). We then structurally and functionally characterise the binding of single-domain nanobodies to the polymerase of the 1918 pandemic influenza virus. Combining these functional and structural data we identify five sites on the polymerase which are sensitive to inhibition by nanobodies. We propose that the binding of nanobodies at these sites either prevents the polymerase from assuming particular functional conformations or interactions with viral or host factors. The polymerase is highly conserved across the influenza A subtypes, suggesting these sites as effective targets for potential influenza antiviral development. Influenza viruses carry their own RNAdependent RNA-polymerase that is highly conserved and a promising anti-viral target. Combining functional and structural data, Keown et al. characterise the inhibitory effect of nanobodies on 1918 pandemic H1N1 influenza strain polymerase complex and identify sensitive sites interfering with polymerase activity in vitro.
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影响因子：
5.4
作者：
Martínez-Alonso M;Hengrung N;Fodor E
通讯作者：
Fodor E
影响因子：
16.8
作者：
Kouba T;Drncová P;Cusack S
通讯作者：
Cusack S
影响因子：
5.4
作者：
Beigel, John H.;Hayden, Frederick G.
通讯作者：
Hayden, Frederick G.
DOI：
10.1038/nrmicro3367
发表时间：
2015-01
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
Eisfeld AJ;Neumann G;Kawaoka Y
通讯作者：
Kawaoka Y
影响因子：
64.8
作者：
Hengrung N;El Omari K;Serna Martin I;Vreede FT;Cusack S;Rambo RP;Vonrhein C;Bricogne G;Stuart DI;Grimes JM;Fodor E
通讯作者：
Fodor E