Complete nucleotide sequence of polyprotein gene 1 and genome organization of turkey coronavirus

Cao, Jian; Wu, Ching Ching; Lin, Tsang Long

doi:10.1016/j.virusres.2008.04.015

Cited by 47 publications

(58 citation statements)

References 43 publications

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“…The biological relevance of these nsps is not clear, but they are believed to be involved in RNA genome replication and transcription. We predicted that there are 8 enzyme domains within the polyprotein 1ab of TCoV, including 3-CLpro, PLP, RdRp, and helicase [20] . Within coronavirus, the in vitro activities of the other four enzymes were recently reported.…”

Section: Discussionmentioning

confidence: 99%

“…TCoV, along with infectious bronchitis virus (IBV) in chickens, is a group 3 coronavirus within the family Coronaviridae . The whole genome of TCoV is 27.7 kb and encodes 13 ORFs [20] . Polyprotein mapping predicted the presence of 14 nsps for pp1ab in which nsp15 shared 40% sequence similarity with nsp15 of SARS-CoV and other coronaviruses and was predicted to be NendoU [20] .…”

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confidence: 99%

“…The whole genome of TCoV is 27.7 kb and encodes 13 ORFs [20] . Polyprotein mapping predicted the presence of 14 nsps for pp1ab in which nsp15 shared 40% sequence similarity with nsp15 of SARS-CoV and other coronaviruses and was predicted to be NendoU [20] . The objective of the present study was to further characterize the enzymatic activity and substrate specificity of the TCoV nsp15.…”

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confidence: 99%

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Turkey Coronavirus Non-Structure Protein NSP15 – An Endoribonuclease

Cao

Lin

2008

Intervirology

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Turkey coronavirus (TCoV) polyprotein was predicted to be cleaved into 15 non-structural proteins (nsp2 to nsp16), but none of these nsps have been characterized. TCoV nsp15 consists of 338 residues and shares 40% sequence similarity to U-specific Nidovirales endoribonuclease (NendoU) of severe acute respiratory syndrome coronavirus. Objective: The purpose of the present study was to characterize TCoV nsp15. Methods: The TCoV nsp15 gene was cloned into pTriEX1 and expressed as a C-terminal His-tagged recombinant protein in BL21 (DE3). The recombinant nsp15 was purified by Ni-NTA resin. Synthetic RNA substrates were used to determine the substrate specificity of the TCoV nsp15. RNA zymography was used to determine the active form of the nsp15. Results: The TCoV nsp15 did not cleave DNA but degraded total cellular RNA. The TCoV nsp15 cleaved single-stranded (ss) RNA at the uridylate site. The TCoV nsp15 cleaved hairpin RNA, pRNA, and double-stranded RNA (dsRNA) of infectious bursal disease virus very slowly, implying that dsRNA is not a good substrate for the TCoV nsp15. No divalent metal ion was required for in vitro enzymatic activity of the TCoV nsp15. The active form of the TCoV nsp15 was a homohexamer and disulfide bond was essential for the enzymatic activity. Conclusion: The TCoV nsp15 is a NendoU but has some characteristics different from other NendoU.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Turkey Coronavirus Non-Structure Protein NSP15 – An Endoribonuclease

Cao

Lin

2008

Intervirology

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“…So far, only North American strains of TCoV (NA TCoV) have been extensively sequenced and characterized at the molecular level (Lin et al, 2004;Cao et al, 2008;Gomaa et al, 2008;Jackwood et al, 2010). These data clearly showed sequence homogeneity between the studied strains with at least 92.4% nucleotide identity for the full-length genomes and 91% amino acid identity for the S proteins.…”

Section: First Full-length Sequences Of the S Gene Of European Isolatmentioning

confidence: 95%

First full-length sequences of the S gene of European isolates reveal further diversity among turkey coronaviruses

et al. 2011

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An increasing incidence of enteric disorders clinically suggestive of the poult enteritis complex has been observed in turkeys in France since 2003. Using a newly designed real-time reverse transcriptase-polymerase chain reaction assay specific for the nucleocapsid (N) gene of infectious bronchitis virus (IBV) and turkey coronaviruses (TCoV), coronaviruses were identified in 37% of the intestinal samples collected from diseased turkey flocks. The full-length spike (S) gene of these viruses was amplified, cloned and sequenced from three samples. The French S sequences shared 98% identity at both the nucleotide and amino acid levels, whereas they were at most 65% and 60% identical with North American (NA) TCoVand at most 50% and 37% identical with IBV at the nucleotide and amino acid levels, respectively. Higher divergence with NA TCoV was observed in the S1-encoding domain. Phylogenetic analysis based on the S gene revealed that the newly detected viruses form a sublineage genetically related with, but significantly different from, NA TCoV. Additionally, the RNAdependent RNA polymerase gene and the N gene, located on the 5? and 3? sides of the S gene in the coronavirus genome, were partially sequenced. Phylogenetic analysis revealed that both the NA TCoV and French TCoV (Fr TCoV) lineages included some IBV relatives, which were however different in the two lineages. This suggested that different recombination events could have played a role in the evolution of the NA and Fr TCoV. The present results provide the first S sequence for a European TCoV. They reveal extensive genetic variation in TCoV and suggest different evolutionary pathways in North America and Europe.

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“…TCoV is an enveloped virus with a liner positive-sense, single-stranded RNA genome with a size of 27 kilobases (Kb). Spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins are four major structure proteins of TCoV Cao et al, 2008). The amino-terminal S1 subunit of S protein forms the globular head of the spike structure on the surface of coronaviruses and contains the receptor binding domain and neutralizing epitopes to determine host tropism and humoral immunity (Godet et al, 1994;Taguchi and Kubo, 1995;He et al, 2004;Ho et al, 2004;Lu et al, 2004;Zhou et al, 2004;Chen et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

A DNA prime-protein boost vaccination strategy targeting turkey coronavirus spike protein fragment containing neutralizing epitope against infectious challenge

Chen

Yoon

et al. 2013

Veterinary Immunology and Immunopathology

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The present study was undertaken to determine immune response and protection efficacy of a spike (S) protein fragment containing neutralizing epitopes (4F/4R) of turkey coronavirus (TCoV) by priming with DNA vaccine and boosting with the recombinant protein from the corresponding DNA vaccine gene segment. Turkeys were vaccinated by priming with either one dose (G1-750DP) or two doses (G3-750DDP) of 750μg DNA vaccine expressing 4F/4R S fragment and boosting with one dose of 200μg 4F/4R S fragment. One dose of 100μg DNA vaccine mixed with polyethyleneimine (PEI) and sodium hyaluronate (HA) followed by one dose of 750μg DNA vaccine and one dose of 200μg 4F/4R S fragment were given to the turkeys in group G2-100DPH. After infectious challenge by TCoV, clinical signs and TCoV detected by immunofluorescence antibody (IFA) assay were observed in less number of turkeys in vaccination groups than that in challenge control groups. TCoV viral RNA loads measured by quantitative real-time reverse transcription-PCR were lower in vaccinated turkeys than those in challenge control turkeys. The turkeys in G3-750DDP produced the highest level of TCoV S protein-specific antibody and virus neutralization (VN) titer. Comparing to the turkeys in G1-750DP, significantly less TCoV were detected by IFA in the turkeys in G2-100DPH receiving an extra dose of 100μg DNA mixed with PEI and HA. The results indicated that DNA-prime protein-boost DNA vaccination regimen targeting TCoV S fragment encompassing neutralizing epitopes induced humoral immune response and partially protected turkeys against infectious challenge by TCoV.

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Complete nucleotide sequence of polyprotein gene 1 and genome organization of turkey coronavirus

Cited by 47 publications

References 43 publications

Turkey Coronavirus Non-Structure Protein NSP15 – An Endoribonuclease

Turkey Coronavirus Non-Structure Protein NSP15 – An Endoribonuclease

First full-length sequences of the S gene of European isolates reveal further diversity among turkey coronaviruses

A DNA prime-protein boost vaccination strategy targeting turkey coronavirus spike protein fragment containing neutralizing epitope against infectious challenge

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