Structure and Function Studies of the Nucleocapsid Protein of Mouse Hepatitis Virus

Masters, Paul S.; Parker, Monica M.; Ricard, Cynthia S.; Duchala, Cynthia S.; Frana, Mark F.; Holmes, Kathryn V.; Sturman, Lawrence S.

doi:10.1007/978-1-4684-5823-7_33

Cited by 16 publications

(14 citation statements)

References 10 publications

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“…The N protein is the major structural protein of CoVs [10][11][12]. The formation of the RNP is important for maintaining the RNA in an ordered conformation that is suitable for viral genome replication and transcription [13][14][15][16]. Previous studies have shown that the CoV N protein is involved in the regulation of cellular processes, such as gene transcription, actin reorganization, host cell cycle progression, and apoptosis [17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Crystal structure-based exploration of the important role of Arg106 in the RNA-binding domain of human coronavirus OC43 nucleocapsid protein

Chen

Yuann

Chang

et al. 2013

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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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 2.0Å. 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.

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Section: Introductionmentioning

confidence: 99%

Crystal structure-based exploration of the important role of Arg106 in the RNA-binding domain of human coronavirus OC43 nucleocapsid protein

Chen

Yuann

Chang

et al. 2013

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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show abstract

“…[10][11][12] Nucleocapsid protein, the major structural protein of CoVs, binds to the viral RNA genome to form the virion core, which leads to the formation of a ribonucleoprotein (RNP) complex or to a long helical nucleocapsid structure. 13,14 The formation of the RNP is important for maintaining the RNA in an ordered conformation suitable for replication and transcription of the viral genome. 13,[15][16][17] Previous studies have shown that the CoV N protein is involved in the regulation of cellular processes such as gene transcription, actin reorganization, host cell cycle progression, and apoptosis.…”

Section: Introductionmentioning

confidence: 99%

Elucidation of the stability and functional regions of the human coronavirus OC43 nucleocapsid protein

et al. 2009

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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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“…6). This is unusual as N proteins are normally highly conserved; alignment of N gene amino acid sequences from five isolates of MHV showed at least 90~ identity (Masters et al, 1990). Nevertheless, variation was mainly clustered in two regions of the N molecule, between positions 204 and 210, and 352 and 359 (Fig.…”

Section: Discussionmentioning

confidence: 94%

“…6). It has been proposed that these two loci represent spacers, which have little sequence specificity but connect conserved domains of the molecule involved in interaction with the RNA genome (Masters et al, 1990). (Masters et al, 1990).…”

Section: Discussionmentioning

confidence: 99%

Analysis of a 9.6 kb sequence from the 3' end of canine coronavirus genomic RNA

Horsburgh

Brierley

Brown

1992

Journal of General Virology

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Structure and Function Studies of the Nucleocapsid Protein of Mouse Hepatitis Virus

Cited by 16 publications

References 10 publications

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

Elucidation of the stability and functional regions of the human coronavirus OC43 nucleocapsid protein

Analysis of a 9.6 kb sequence from the 3' end of canine coronavirus genomic RNA

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