Zuwała Kaja, Golda Anna, Kabala Wojciech, Burmistrz Michał, Zdzalik Michal, Nowak Paulina, Kedracka-Krok Sylwia, Zarebski Mirosław, Dobrucki Jerzy, Florek Dominik, Zeglen Sławomir, Wojarski Jacek, Potempa Jan, Dubin Grzegorz, Pyrc Krzysztof
Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387, Krakow, Poland.
Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387, Krakow, Poland; Malopolska Centre of Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387, Krakow, Poland.
PLoS One. 2015 Feb 20;10(2):e0117833. doi: 10.1371/journal.pone.0117833. eCollection 2015.
Human coronavirus (HCoV) NL63 was first described in 2004 and is associated with respiratory tract disease of varying severity. At the genetic and structural level, HCoV-NL63 is similar to other members of the Coronavirinae subfamily, especially human coronavirus 229E (HCoV-229E). Detailed analysis, however, reveals several unique features of the pathogen. The coronaviral nucleocapsid protein is abundantly present in infected cells. It is a multi-domain, multi-functional protein important for viral replication and a number of cellular processes. The aim of the present study was to characterize the HCoV-NL63 nucleocapsid protein. Biochemical analyses revealed that the protein shares characteristics with homologous proteins encoded in other coronaviral genomes, with the N-terminal domain responsible for nucleic acid binding and the C-terminal domain involved in protein oligomerization. Surprisingly, analysis of the subcellular localization of the N protein of HCoV-NL63 revealed that, differently than homologous proteins from other coronaviral species except for SARS-CoV, it is not present in the nucleus of infected or transfected cells. Furthermore, no significant alteration in cell cycle progression in cells expressing the protein was observed. This is in stark contrast with results obtained for other coronaviruses, except for the SARS-CoV.
人类冠状病毒(HCoV)NL63于2004年首次被描述,与不同严重程度的呼吸道疾病有关。在基因和结构水平上,HCoV-NL63与冠状病毒亚科的其他成员相似,尤其是人类冠状病毒229E(HCoV-229E)。然而,详细分析揭示了该病原体的几个独特特征。冠状病毒核衣壳蛋白大量存在于受感染细胞中。它是一种多结构域、多功能蛋白,对病毒复制和许多细胞过程都很重要。本研究的目的是对HCoV-NL63核衣壳蛋白进行表征。生化分析表明,该蛋白与其他冠状病毒基因组中编码的同源蛋白具有共同特征,其N端结构域负责核酸结合,C端结构域参与蛋白寡聚化。令人惊讶的是,对HCoV-NL63 N蛋白亚细胞定位的分析表明,与除SARS-CoV外的其他冠状病毒物种的同源蛋白不同,它不存在于受感染或转染细胞的细胞核中。此外,在表达该蛋白的细胞中未观察到细胞周期进程的显著改变。这与除SARS-CoV外的其他冠状病毒的结果形成鲜明对比。