Journal
PROTEIN SCIENCE
Volume 18, Issue 11, Pages 2209-2218Publisher
WILEY
DOI: 10.1002/pro.225
Keywords
human coronavirus; OC43 strain; nucleocapsid protein; Ribonucleocapsid; stability; RNA-binding regions; oligomerization domain
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Funding
- NCS [97-2311-B-005-003-MY3]
- Grants-in-Aid for Scientific Research [24570176] Funding Source: KAKEN
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Human coronavirus OC43 (HCoV-OC43) is one of the causes of the common cold in human during seasons of cold weather. The primary function of the HCoV-OC43 nucleocapsid protein (N protein) is to recognize viral genomic RNA, which leads to ribonucleocapsid formation. Here, we characterized the stability and identified the functional regions of the recombinant HCoV-OC43 N protein. Circular dichroism and fluorescence measurements revealed that the HCoV-OC43 N protein is more highly ordered and stabler than the SARS-CoV N protein previously studied. Surface plasmon resonance (SPR) experiments showed that the affinity of HCoV-OC43 N protein for RNA was approximately fivefold higher than that of N protein for DNA. Moreover, we found that the HCoV-OC43 N protein contains three RNA-binding regions in its N-terminal region (residues 1-173) and central-linker region (residues 174-232 and 233-300). The binding affinities of the truncated N proteins and RNA follow the order: residues 1-173-residues 233-300 > residues 174-232. SPR experiments demonstrated that the C-terminal region (residues 301-448) of HCoV-OC43 N protein lacks RNA-binding activity, while crosslinking and gel filtration analyses revealed that the C-terminal region is mainly involved in the oligomerization of the HCoV-OC43 N protein. This study may benefit the understanding of the mechanism of HCoV-OC43 nucleocapsid formation.
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