Crystal structure-based exploration of the important role of Arg106 in the RNA-binding domain of human coronavirus OC43 nucleocapsid protein.
Crystal structure-based exploration of the important role of Arg106 in the RNA-binding domain of human coronavirus OC43 nucleocapsid protein.
复制标题
DOI：
10.1016/j.bbapap.2013.03.003
复制
发表时间：
2013-06
影响因子：
3.2
通讯作者：
Hou, Ming-Hon
中科院分区：
文献类型：
--
作者：
Chen, I. -Jung;Yuann, Jeu-Ming P.;Chang, Yu-Ming;Lin, Shing-Yen;Zhao, Jincun;Perlman, Stanley;Shen, Yo-Yu;Huang, Tai-Huang;Hou, Ming-Hon
人冠状病毒OC 43（HCoV-OC 43）是一种常见的感冒病原体。核衣壳（N）蛋白是CoV的主要结构蛋白，其与病毒RNA基因组结合以形成病毒体核心并导致核糖核蛋白（RNP）复合物的形成。我们已经解析了HCoV-OC 43 N蛋白（N-NTD）N-末端结构域（残基58至195）的晶体结构，分辨率为2.0 nm。HCoV-OC 43 N-NTD是一种单结构域蛋白，由一个五链β折叠核心和一个长的延伸环组成，与其他冠状病毒的N-NTD结构相似。HCoV-OC 43 N-NTD的带正电荷的环含有结构上保守的带正电荷的残基R106。为了评估R106在RNA结合中的作用，我们进行了一系列定点诱变实验和对接模拟来表征R106与RNA之间的相互作用。结果表明，R106在N蛋白与RNA的相互作用中起重要作用。此外，我们发现，在用编码突变体（R106 A）N蛋白的质粒转染并感染病毒的细胞中，与用野生型N蛋白转染的细胞相比，基质蛋白基因的水平降低了7倍。这一发现表明R106通过增强N蛋白与病毒RNA的结合在病毒复制中起关键作用。结果还表明，N蛋白/RNA相互作用的强度对HCoV-OC 43复制至关重要。HCoV-OC 43 N蛋白的NTD负责RNA结合。N-NTD由一个五股β-折叠核心和一个长的延伸环组成。R106在N蛋白与RNA的相互作用中起重要作用。R106在病毒复制中起着关键作用。
Human coronavirus OC43 (HCoV-OC43) is a causative agent of the common cold. The nucleocapsid (N) protein, which is a major structural protein of CoVs, binds to the viral RNA genome to form the virion core and results in the formation of the ribonucleoprotein (RNP) complex. We have solved the crystal structure of the N-terminal domain of HCoV-OC43 N protein (N-NTD) (residues 58 to 195) to a resolution of 2.0 Å. The HCoV-OC43 N-NTD is a single domain protein composed of a five-stranded β-sheet core and a long extended loop, similar to that observed in the structures of N-NTDs from other coronaviruses. The positively charged loop of the HCoV-OC43 N-NTD contains a structurally well-conserved positively charged residue, R106. To assess the role of R106 in RNA binding, we undertook a series of site-directed mutagenesis experiments and docking simulations to characterize the interaction between R106 and RNA. The results show that R106 plays an important role in the interaction between the N protein and RNA. In addition, we showed that, in cells transfected with plasmids that encoded the mutant (R106A) N protein and infected with virus, the level of the matrix protein gene was decreased by 7-fold compared to cells that were transfected with the wild-type N protein. This finding suggests that R106, by enhancing binding of the N protein to viral RNA plays a critical role in the viral replication. The results also indicate that the strength of N protein/RNA interactions is critical for HCoV-OC43 replication. The NTD of the HCoV-OC43 N protein is responsible for RNA-binding. The N-NTD is composed of a five-stranded β-sheet core and a long extended loop. R106 plays an important role in the interaction between the N protein and RNA. R106 plays a critical role in the virus replication.
登录
查看更多内容
DOI：
10.1016/j.str.2005.08.021
发表时间：
2005-12
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Fan H;Ooi A;Tan YW;Wang S;Fang S;Liu DX;Lescar J
通讯作者：
Lescar J
DOI：
10.1128/cdli.12.11.1317-1321.2005
发表时间：
2005-11-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Chan, KH;Cheng, VCC;Peiris, JSM
通讯作者：
Peiris, JSM
影响因子：
2.9
作者：
Huang, QL;Yu, LP;Olejniczak, ET
通讯作者：
Olejniczak, ET
影响因子：
5.4
作者：
Kopecky-Bromberg, Sarah A.;Martinez-Sobrido, Luis;Palese, Peter
通讯作者：
Palese, Peter
影响因子：
3.1
作者：
Liang FY;Lin LC;Ying TH;Yao CW;Tang TK;Chen YW;Hou MH
通讯作者：
Hou MH