The Nucleocapsid Protein of Coronavirus Infectious Bronchitis Virus: Crystal Structure of Its N-Terminal Domain and Multimerization Properties

Fan, Haijun; Ooi, A.; Tan, Yong Kiam; Wang, Sifang; Fang, Shouguo; Liu, Ding Xiang; Lescar, Julien

doi:10.1016/j.str.2005.08.021

Cited by 128 publications

(187 citation statements)

References 43 publications

(47 reference statements)

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“…In this study, we report the crystal structure of the dimerization domain of SARS-CoV N protein consisting of residues 270 -370. Consistent with biochemical studies showing that this domain mediates dimer formation of the N protein (17,23), the structure consists of a dimer with extensive interactions between subunits, suggesting that the N protein is not stable in the monomeric form and that the dimeric structure represents the functional unit of the N protein. Furthermore, the dimerization domain of SARS-CoV N protein shows a similar fold to that of the N protein of porcine reproductive and respiratory syndrome virus (PRRSV), a member of Arteriviridae.…”

supporting

confidence: 76%

Crystal Structure of the Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein Dimerization Domain Reveals Evolutionary Linkage between Corona- and Arteriviridae

Oldham

Zhang

et al. 2006

Journal of Biological Chemistry

127

156

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The causative agent of severe acute respiratory syndrome (SARS) is the SARS-associated coronavirus, SARS-CoV. The nucleocapsid (N) protein plays an essential role in SARS-CoV genome packaging and virion assembly. We have previously shown that SARS-CoV N protein forms a dimer in solution through its C-terminal domain. In this study, the crystal structure of the dimerization domain, consisting of residues 270 -370, is determined to 1.75 Å resolution. The structure shows a dimer with extensive interactions between the two subunits, suggesting that the dimeric form of the N protein is the functional unit in vivo. Although lacking significant sequence similarity, the dimerization domain of SARS-CoV N protein has a fold similar to that of the nucleocapsid protein of the porcine reproductive and respiratory syndrome virus. This finding provides structural evidence of the evolutionary link between Coronaviridae and Arteriviridae, suggesting that the N proteins of both viruses have a common origin.Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses that infect a variety of mammals and birds. Although previously identified human coronaviruses cause only mild respiratory infections, in the 2003 outbreak of severe acute respiratory syndrome (SARS), 3 a disease caused by a new type of coronavirus (SARS-CoV), there were more than 8,000 cases resulting in 774 deaths (ϳ10% mortality). Phylogenetic analysis suggests that SARSCoV diverged early from group 2 coronaviruses and has evolved independently for a long period of time (1).The coronavirus genome, containing ϳ30,000 bases, is the largest among positive-sense RNA viruses (2, 3). It encodes non-structural proteins including the RNA polymerase and helicase, as well as the spike (S), envelope (E), membrane (M), and nucleocapsid (N) structural proteins. The coronavirus virion is about 120 nm in diameter and consists of a lipid envelope containing three or four anchored glycoproteins and a helical ribonucleoprotein core (4). The surface projections forming the crown-like structure observed via electron microscopy are made up of the S protein, which is responsible for receptor recognition and membrane fusion (5-9). The integral membrane proteins M and E are essential for virus budding. When co-expressed in animal cells, the M and E proteins are sufficient to form virus-like particles (10). The N protein interacts with the viral genome to form the ribonucleoprotein core and has been shown to be involved in viral RNA synthesis, transcriptional regulation of genomic RNA, translation of viral proteins, and budding (2,11,12).Coronaviruses are related to arteriviruses by their similar genome organizations and viral replication mechanisms (3, 13). Recently, Coronaviridae and Arteriviridae were united to form the new order Nidovirales. The name of the order comes from a property common to both viruses, a nested set of subgenomic mRNAs for structural protein expression (Latin nidus, meaning nest). The replicase genes of arteriviruses and coronaviruses are thought to ha...

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supporting

confidence: 76%

Crystal Structure of the Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein Dimerization Domain Reveals Evolutionary Linkage between Corona- and Arteriviridae

Oldham

Zhang

et al. 2006

Journal of Biological Chemistry

127

156

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“…Coronaviruses are considered to have a helical nucleocapsid (Davies et al, 1981;Sturman et al, 1980). The exact organization of this nucleocapsid is not known, but some models have been proposed (Chen et al, 2007;Fan et al, 2005). The cryo-EM of SARS-CoV showed an amorphous core with the same elongated shape as that of the whole virion.…”

Section: Comparison With Other Nidovirusesmentioning

confidence: 99%

“…Although it is not known exactly how the coronavirus N protein is organized into a helical nucleocapsid (Davies et al, 1981;Sturman et al, 1980), several possible models have been proposed (Chen et al, 2007;Fan et al, 2005;Jayaram et al, 2006). In some of these models, the RNA forms the centre of a helix formed by the N proteins, while others have proposed that the RNA is wound around the outside of a helical N protein core.…”

mentioning

confidence: 99%

Cryo-electron tomography of porcine reproductive and respiratory syndrome virus: organization of the nucleocapsid

Spilman

Welbon

Nelson

et al. 2009

Journal of General Virology

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Porcine reproductive and respiratory virus (PRRSV) is an enveloped positive-sense RNA virus of the family Arteriviridae that causes severe and persistent disease in pigs worldwide. The PRRSV virion consists of a lipid envelope that contains several envelope proteins surrounding a nucleocapsid core that encapsidates the RNA genome. To provide a better understanding of the structure and assembly of PRRSV, we have carried out cryo-electron microscopy and tomographic reconstruction of virions grown in MARC-145 cells. The virions are pleomorphic, round to egg-shaped particles with an average diameter of 58 nm. The particles display a smooth outer surface with only a few protruding features, presumably corresponding to the envelope protein complexes. The virions contain a double-layered, hollow core with an average diameter of 39 nm, which is separated from the envelope by a 2-3 nm gap. Analysis of the three-dimensional structure suggests that the core is composed of a double-layered chain of nucleocapsid proteins bundled into a hollow ball.

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“…Previous studies have shown that full-length CoV N proteins can form high-order oligomers, and the C-terminal domains of the CoV N proteins are responsible for oligomerization [30,[33][34][35][36][37][38]. Crystal structures of the C-terminal domains of CoV N proteins have been published and suggest that the basic building block for coronavirus nucleocapsid formation is the dimeric assembly of the N protein [34,[39][40][41]. Luo et al revealed that the CoV N protein might combine with viral genomic RNA to generate higher-order oligomers, which could trigger the formation of the long nucleocapsid structure [32].…”

Section: Introductionmentioning

confidence: 99%

Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein

Lin

Wang³

et al. 2012

FEBS Letters

102

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a b s t r a c tThe coronavirus (CoV) N protein oligomerizes via its carboxyl terminus. However, the oligomerization mechanism of the C-terminal domains (CTD) of CoV N proteins remains unclear. Based on the protein disorder prediction system, a comprehensive series of HCoV-229E N protein mutants with truncated CTD was generated and systematically investigated by biophysical and biochemical analyses to clarify the role of the C-terminal tail of the HCoV-229E N protein in oligomerization. These results indicate that the last C-terminal tail plays an important role in dimer-dimer association. The C-terminal tail peptide is able to interfere with the oligomerization of the CTD of HCoV-229E N protein and performs the inhibitory effect on viral titre of HCoV-229E. This study may assist the development of anti-viral drugs against HCoV. Structured summary of protein interactions:N and C-terminal tail peptide bind by cosedimentation in solution (View interaction) N and N bind by cosedimentation in solution (View Interaction: 1,2,3,4,5,6,7,8,9,10,11,12) C-terminal tail peptide and N bind by fluorescence technology (View interaction) N and N bind by cross-linking study (View interaction) N and N bind by cross-linking study (View Interaction: 1,2,3,4) Crown

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The Nucleocapsid Protein of Coronavirus Infectious Bronchitis Virus: Crystal Structure of Its N-Terminal Domain and Multimerization Properties

Cited by 128 publications

References 43 publications

Crystal Structure of the Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein Dimerization Domain Reveals Evolutionary Linkage between Corona- and Arteriviridae

Crystal Structure of the Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein Dimerization Domain Reveals Evolutionary Linkage between Corona- and Arteriviridae

Cryo-electron tomography of porcine reproductive and respiratory syndrome virus: organization of the nucleocapsid

Oligomerization of the carboxyl terminal domain of the human coronavirus 229E nucleocapsid protein

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