Molecular Basis of Transmissible Gastroenteritis Virus Epidemiology

Enjuanes, Luis; Zeijst, B. A. M. Van Der

doi:10.1007/978-1-4899-1531-3_16

Cited by 81 publications

(103 citation statements)

References 184 publications

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“…RT-PCR of Leader-Containing Transcripts. Leader-containing amplicons were obtained from WT and icSARS-CoV-infected cells by using primers at the 3Ј end of the genome (5Ј-tttttttttttttttttttttgtcattctcctaagaagc 29710 -3Ј) or in the X5 or X3 ORFs (5Ј-ttaattaattaatttgttcgtttatttaaaacaaca 28091Ј -3Ј, 5Ј-ttaattaattatggataatctaactccataggttct 27238 -3Ј) and in the SARS leader RNA sequence at the 5Ј end of the genome (5Ј-aaagccaaccaacctcgatc-3Ј; nucleotides [26][27][28][29][30][31][32][33][34][35]. Leader-containing amplicons were subcloned into TopoII vectors and sequenced.…”

Section: Methodsmentioning

confidence: 99%

Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Yount

Curtis

Fritz

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

349

400

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Section: Methodsmentioning

confidence: 99%

Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Yount

Curtis

Fritz

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

349

400

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“…In addition to the viral mRNAs, full-length and subgenome-length negativestrand RNAs are implicated in mRNA synthesis (4,26,40,42,43). Another unique feature of coronavirus replication is the high RNA recombination frequencies associated with infection (6,25,26).The large size of the coronavirus genome, coupled with the inability to clone portions of the polymerase gene in microbial vectors, has hampered the ability to perform precise manipulations and reverse genetics in members of the Coronaviridae (17,18,26,44). Recently these problems were overcome when a full-length cDNA of TGEV was stably cloned in bacterial artificial chromosome (BAC) vectors (3).…”

mentioning

confidence: 99%

Strategy for Systematic Assembly of Large RNA and DNA Genomes: Transmissible Gastroenteritis Virus Model

Yount

Curtis

Baric

2000

J Virol

244

284

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A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The ϳ30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineered with unique flanking interconnecting junctions which determine a precise systematic assembly with only the adjacent cDNA subclones, resulting in an intact TGEV cDNA construct of ϳ28.5 kb in length. Transcripts derived from the full-length TGEV construct were infectious, and progeny virions were serially passaged in permissive host cells. Viral antigen production and subgenomic mRNA synthesis were evident during infection and throughout passage. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar plaque morphology in permissive host cells. Host range phenotypes of the molecularly cloned and wild-type viruses were similar in cells of swine and feline origin. The recombinant viruses were sequenced across the unique interconnecting junctions, conclusively demonstrating the marker mutations and restriction sites that were engineered into the component clones. Full-length infectious constructs of TGEV will permit the precise genetic modification of the coronavirus genome. The method that we have designed to generate an infectious cDNA construct of TGEV could theoretically be used to precisely reconstruct microbial or eukaryotic genomes approaching several million base pairs in length.Molecular genetic analysis of the structure and function of RNA virus genomes has been profoundly advanced by the availability of full-length cDNA clones, the source of infectious RNA transcripts that replicate efficiently when introduced into permissive cell lines (2, 9). Recombinant DNA technology has allowed the isolation of infectious cDNA clones from a number of positive-stranded RNA viruses, including picornaviruses, caliciviruses, alphaviruses, flaviviruses, and arteriviruses, whose RNA genomes range in size from ϳ7 to 15 kb in length (1,13,32,34,35,47,48,54). The availability of these clones has enhanced our understanding of the molecular mechanisms of viral replication and pathogenesis and resulted in new approaches for heterologous gene expression and vaccine development.The order Nidovirales includes mammalian positive-polarity single-stranded RNA viruses in the arterivirus and coronavirus families (10, 16). The Coronaviridae family includes the Coronavirus and Torovirus genera (10, 46). Despite significant size differences (ϳ13 to 32 kb), the polycistronic genome organization and regulation of gene expression from a nested set of subgenomic mRNAs are similar for all members of the order (16, 4...

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“…TGEV replicates in enterocytes and provokes villous atrophy, is closely related to the human respiratory coronavirus HCoV-229E (9), and can also infect the respiratory tract. Moreover, some variant strains of TGEV, such as the porcine respiratory coronavirus (PRCoV), have lost their intestinal tropism (11,18). The Purdue-115 strain (10) and the Miller strain (34) of TGEV have been shown to induce apoptosis in cell lines expressing the porcine aminopeptidase N (APN), which is a receptor for the virus (4).…”

mentioning

confidence: 99%

The Viral Nucleocapsid Protein of Transmissible Gastroenteritis Coronavirus (TGEV) Is Cleaved by Caspase-6 and -7 during TGEV-Induced Apoptosis

Éléouët

Slee

Saurini

et al. 2000

J Virol

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The transmissible gastroenteritis coronavirus (TGEV), like many other viruses, exerts much of its cytopathic effect through the induction of apoptosis of its host cell. Apoptosis is coordinated by a family of cysteine proteases, called caspases, that are activated during apoptosis and participate in dismantling the cell by cleaving key structural and regulatory proteins. We have explored the caspase activation events that are initiated upon infection of the human rectal tumor cell line HRT18 with TGEV. We show that TGEV infection results in the activation of caspase-3, -6, -7, -8, and -9 and cleavage of the caspase substrates eIF4GI, gelsolin, and ␣-fodrin. Surprisingly, the TGEV nucleoprotein (N) underwent proteolysis in parallel with the activation of caspases within the host cell. Cleavage of the N protein was inhibited by cell-permeative caspase inhibitors, suggesting that this viral structural protein is a target for host cell caspases. We show that the TGEV nucleoprotein is a substrate for both caspase-6 and -7, and using site-directed mutagenesis, we have mapped the cleavage site to VVPD 359 2. These data demonstrate that viral proteins can be targeted for destruction by the host cell death machinery.

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Molecular Basis of Transmissible Gastroenteritis Virus Epidemiology

Cited by 81 publications

References 184 publications

Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Strategy for Systematic Assembly of Large RNA and DNA Genomes: Transmissible Gastroenteritis Virus Model

The Viral Nucleocapsid Protein of Transmissible Gastroenteritis Coronavirus (TGEV) Is Cleaved by Caspase-6 and -7 during TGEV-Induced Apoptosis

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