Fe-S cofactors in the SARS-CoV-2 RNA-dependent RNA polymerase are potential antiviral targets.
Fe-S cofactors in the SARS-CoV-2 RNA-dependent RNA polymerase are potential antiviral targets.
复制标题
DOI：
10.1126/science.abi5224
复制
发表时间：
2021-07-09
期刊：
影响因子：
--
通讯作者：
Rouault TA
中科院分区：
文献类型：
--
作者：
Maio N;Lafont BAP;Sil D;Li Y;Bollinger JM Jr;Krebs C;Pierson TC;Linehan WM;Rouault TA
铁硫团簇是参与代谢和电子转移的蛋白质的重要辅助因子，但有时也在参与DNA转录和复制的酶中发现。这种酶的体外表达可能导致错误的簇组装和对功能酶的组成的混淆。通过仔细的缺氧净化方案，Maio等人发现，严重急性呼吸综合征冠状病毒2 RNA依赖的RNA聚合酶在先前观察到的两个结合锌离子的位点上含有两个铁硫簇。连接半胱氨酸残基的突变导致聚合酶活性的丧失。含锌酶的活性丧失程度较轻。用氮氧化物药物TEMPOL处理导致细胞培养中的簇降解、酶抑制和病毒复制抑制。靶向RNA依赖的RNA聚合酶Fe-S簇体外抑制SARS-CoV-2感染。严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）是COVID-19的致病因子，它使用RNA依赖性RNA聚合酶（RdRp）进行基因组复制和基因转录。我们发现RdRp的催化亚基nsp12连接了两个铁硫金属辅因子，这些辅因子在RdRp配合物的可用低温电子显微镜结构中被模拟为锌中心。这些金属结合位点对于复制和与病毒解旋酶的相互作用是必不可少的。稳定的氮氧化物TEMPOL氧化团簇导致其分解，有效抑制RdRp，并阻断了细胞培养中SARS-CoV-2的复制。因此，这些铁硫簇是SARS-CoV-2 RdRp的辅助因子，也是COVID-19治疗的靶点。
Iron–sulfur clusters are important cofactors for proteins involved in metabolism and electron transfer but are also sometimes found in enzymes involved in transcription and replication of DNA. In vitro expression of such enzymes can result in faulty cluster assembly and confusion about the composition of the functional enzyme. Using a careful anoxic purification scheme, Maio et al. found that the severe acute respiratory syndrome coronavirus 2 RNA–dependent RNA polymerase contains two iron–sulfur clusters at two sites previously observed to bind zinc ions. Mutation of the ligating cysteine residues resulted in loss of polymerase activity. A less severe loss of activity was seen in the zinc-containing enzyme. Treatment with the nitroxide drug TEMPOL resulted in degradation of the clusters, enzyme inhibition, and inhibition of viral replication in cell culture. Science, abi5224, this issue p. 236 Targeting RNA-dependent RNA polymerase Fe-S clusters inhibits SARS-CoV-2 infection in vitro. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of COVID-19, uses an RNA-dependent RNA polymerase (RdRp) for the replication of its genome and the transcription of its genes. We found that the catalytic subunit of the RdRp, nsp12, ligates two iron-sulfur metal cofactors in sites that were modeled as zinc centers in the available cryo–electron microscopy structures of the RdRp complex. These metal binding sites are essential for replication and for interaction with the viral helicase. Oxidation of the clusters by the stable nitroxide TEMPOL caused their disassembly, potently inhibited the RdRp, and blocked SARS-CoV-2 replication in cell culture. These iron-sulfur clusters thus serve as cofactors for the SARS-CoV-2 RdRp and are targets for therapy of COVID-19.
登录
查看更多内容
影响因子：
168.9
作者：
Chan, Jasper Fuk-Woo;Yuan, Shuofeng;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
影响因子：
2.9
作者：
CUNNINGHAM, RP;ASAHARA, H;EMPTAGE, MH
通讯作者：
EMPTAGE, MH
DOI：
10.1074/jbc.m112.368951
发表时间：
2012-07-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kilkenny ML;De Piccoli G;Perera RL;Labib K;Pellegrini L
通讯作者：
Pellegrini L
影响因子：
64.8
作者：
Gordon, David E.;Jang, Gwendolyn M.;Krogan, Nevan J.
通讯作者：
Krogan, Nevan J.
影响因子：
16.8
作者：
Klinge, Sebastian;Hirst, Judy;Maman, Joseph D;Krude, Torsten;Pellegrini, Luca
通讯作者：
Pellegrini, Luca