Nucleotide Sequence of Porcine Circovirus Associated with Postweaning Multisystemic Wasting Syndrome in Pigs

Hamel, André; Lin, Lihua; Nayar, Gopi

doi:10.1128/jvi.72.6.5262-5267.1998

Cited by 384 publications

(265 citation statements)

References 18 publications

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“…Circoviruses are the smallest autonomously replicating DNA viruses, and they show a typical ambisense genomic structure [9,10]. Circoviruses have a non-enveloped, circular, single-stranded DNA genome consisting of two major open reading frames, ORF1 and ORF2, which code the replicase (Rep) and the capsid protein (Cap), respectively [11,12].…”

Section: Introductionmentioning

confidence: 99%

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Mou

Wang

Pan

et al. 2019

Viruses

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Porcine circovirus type 3 (PCV3) contains two major open reading frames (ORFs) and the ORF2 gene encodes the major structural capsid protein. In this study, nuclear localization of ORF2 was demonstrated by fluorescence observation and subcellular fractionation assays in ORF2-transfected PK-15 cells. The subcellular localization of truncated ORF2 indicated that the 38 N-terminal amino acids were responsible for the nuclear localization of ORF2. The truncated and site-directed mutagenesis of this domain were constructed, and the results demonstrated that the basic amino acid residues at positions 8-32 were essential for the strict nuclear localization. The basic motifs 8 RRR-R-RRR 16 and 16 RRRHRRR 22 were further shown to be the key functional nucleolar localization signals that guide PCV3 ORF2 into nucleoli. Furthermore, sequence analysis showed that the amino acids of PCV3 nuclear localization signals were highly conserved. Overall, this study provides insight into the biological and functional characteristics of the PCV3 ORF2 protein.case of PCVs, the N-terminus of ORF2 is rich in basic amino acids and displays nuclear localization signals (NLSs) [14][15][16]. The NLS is a short stretch of amino acids that mediates the transport of nuclear proteins into the nucleus. NLS motifs play a key role in this mechanism. NLS sequences are often composed of basic amino acids and can be classified as either monopartite or bipartite motifs [17,18]. The nuclear targeting of PCV2 ORF2 is directed by the bipartite motifs situated at the N-terminus of the proteins [14,15]. NLSs of PCV1 ORF2 show high homologies to classical monopartite or bipartite NLS, which is essential for the complete nuclear import of PCV1 ORF2. The N-terminus of PCV1 and PCV2 ORF2 contain several conserved basic amino acid stretches [15,19], and shares 70.7% in nucleotide identity and 82.9% in amino acid similarity.Verification of NLS and demonstration of intracellular distribution facilitate understanding of viral protein function. The amino acids of PCV3 ORF2 are markedly different from those of the other PCVs. This led us to study the functional motifs in nuclear targeting of PCV3 ORF2. In this study, a series of recombinant plasmids expressing PCV3 ORF2 fused to EGFP were constructed, and we identified the main motifs of the NLSs mediating nuclear localization of PCV3 ORF2.

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Section: Introductionmentioning

confidence: 99%

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Mou

Wang

Pan

et al. 2019

Viruses

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“…Other common lesions described for PMWS include interstitial pneumonia, periportal to diffuse hepatitis, and interstitial nephritis (Clark, 1997;Rosell et al, 1999). Isolation of virus from tissues of affected pigs led to the identification of a porcine circovirus type 2 (PCV2), considered to be of etiological importance in PMWS Ellis et al, 1998;Hamel et al, 1998;Meehan et al, 1998;Morozov et al, 1998). Porcine circovirus type 1 (PCV1) was originally identified as a contaminant of porcine kidney cell cultures (PK15 ATCC CCL-33) (Tischer et al, 1982) and is nonpathogenic to pigs (Allan et al, 1995;Tischer et al, 1982).…”

Section: Introductionmentioning

confidence: 99%

“…This discovery promoted the use of RNAi for specifically inhibiting gene expression and replication of infectious viruses. Over the past years, RNAi approaches have shown to be effective against a variety of viruses in cell culture, among these are prominent viruses such as human immunodeficiency virus 1 (Capodici et al, 2002), influenza virus (Ge et al, 2003), poliovirus (Gitlin et al, 2002), severe acute respiratory syndrome virus (Wang et al, 2004), herpes virus (Jia and Sun, 2003), human papillomavirus (Hamel et al, 1998), dengue virus (Caplen et al, 2002), and West Nile virus (McCown et al, 2003). Synthetic siRNAs as well as shRNAs transcribed in vivo from DNA templates have been shown to be potent inhibitors of gene expression in adult mice (McCaffrey et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of porcine circovirus type 2 replication in mice by RNA interference

et al. 2006

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Porcine circovirus type 2 (PCV2) is the primary causative agent of an emerging swine disease, postweaning multisystemic wasting syndrome (PMWS) for which no antiviral treatment is available. To exploit the possibility of using RNA interference (RNAi) as a therapeutic approach against the disease, plasmid-borne short hairpin RNAs (shRNAs) were generated to target the PCV2 genome. Transfection of these shRNAs into cultured PK15 cells caused a significant reduction in viral RNA production that was accompanied by inhibiting viral DNA replication and protein synthesis in infected cells. The effect was further tested in vivo in a mouse model that has been developed for PCV2 infection. Mice injected with shRNA before PCV2 infection showed substantially decreased microscopic lesions in inguinal lymph nodes compared to controls. In situ hybridization and immunohistochemical analyses showed that shRNA caused a significant inhibition in the level of viral DNA and protein synthesis detected in the lymph nodes of the treated mice relative to the controls. Taken together, these results indicate that shRNAs are capable of inhibiting PCV2 infection in vitro as well as in vivo and thus may constitute an effective therapeutic strategy for PCV2 infection.

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“…First identified in western Canada in 1991 (Clark, 1997), this disease, known as post-weaning multisystemic wasting syndrome (PMWS), was subsequently confirmed and described in Spain (Segalès et al, 1997), France , the United States (Allan et al, 1998a), Ireland and Denmark . Porcine circovirus type 2 (PCV2) was detected in the tissues of affected animals (Allan et al, 1998b;Ellis et al, 1998;Hamel et al, 1998;Meehan et al, 1998;Morozov et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome

Blanchard

Mahé

Cariolet

et al. 2003

Veterinary Microbiology

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Porcine circovirus type 2 (PCV2) plays a crucial role in the pathogenesis of post-weaning multisystemic wasting syndrome (PMWS) in swine. As PCV2 displays significant homology with PCV1 (a non-pathogenic virus) at the nucleotide and amino-acid level, a discriminative antigen is needed for specific serological diagnosis. The ORF2-encoded capsid protein from PCV2 was used to develop an indirect enzyme-linked immunosorbent assay (ELISA). GST-fused capsid protein from PCV2 and GST alone (both expressed in recombinant baculovirus-infected cells) were used as antigens for serodiagnosis. The specificity of the ELISA for detection of PCV2 antibodies was demonstrated in sera from pigs experimentally infected with PCV1, PCV2 and other swine viruses. The semi-quantitative nature of the test was evaluated versus an immunoperoxidase monolayer assay (IPMA). The ELISA was performed on 322 sera from pigs in eight Brittany herds and compared with IPMA. The sensitivity (98.2%) and specificity (94.5%) of this test were considered suitable for individual serological detection. High PCV2 seroprevalence was found in sows and pigs at the end of the growth phase (18-19 weeks) in all eight herds. The seroprevalence in piglets (11-17 weeks) was statistically correlated with clinical symptoms of PMWS (93% in affected versus 54%, in non-affected farms). A cohort study performed in PMWS-free farms showed that 57% of piglets exhibited active seroconversion after 13 weeks, indicating that PCV2 infection occurred earlier in PMWS-affected piglets.

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Nucleotide Sequence of Porcine Circovirus Associated with Postweaning Multisystemic Wasting Syndrome in Pigs

Cited by 384 publications

References 18 publications

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Inhibition of porcine circovirus type 2 replication in mice by RNA interference

An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome

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