Processing and evolution of the N-terminal region of the arterivirus replicase ORF1a protein: identification of two papainlike cysteine proteases

Boon, Johan A. den; Faaberg, Kay S.; Meulenberg, J.J.M.; Wassenaar, Alfred L. M.; Plagemann, Peter G.W.; Gorbalenya, Alexander E.; Snijder, Eric J.

doi:10.1128/jvi.69.7.4500-4505.1995

Cited by 172 publications

(137 citation statements)

References 30 publications

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“…Translation of ORF1a yields the pp1a polyprotein. ORF1b is expressed through a ribosomal frame shift, leading to the formation of a large pp1ab polyprotein (den Boon et al, 1995;Meulenberg et al, 1993). The pp1a and pp1ab polyproteins are processed by viral proteases to release 14 non-structural proteins, which include four proteases (NSP1␣, NSP1␤, NSP2 and NSP4), the RNA-dependent RNA polymerase (NSP9), a helicase (NSP10) and an endonuclease (NSP11) (den Boon et al, 1995;Snijder and Meulenberg, 1998;van Aken et al, 2006;Ziebuhr et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

The structural biology of PRRSV

2010

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Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped, positive-sense single-stranded RNA virus belonging to the Arteriviridae family. Arteriviruses and coronaviruses are grouped together in the order Nidovirales, based on similarities in genome organization and expression strategy. Over the past decade, crystal structures of several viral proteins, electron microscopic studies of the virion, as well as biochemical and in vivo studies on protein-protein interactions have led to a greatly increased understanding of PRRSV structural biology. At this point, crystal structures are available for the viral proteases NSP1α, NSP1β and NSP4 and the nucleocapsid protein, N. The NSP1α and NSP1β structures have revealed additional non-protease domains that may be involved in modulation of host functions. The N protein forms a dimer with a novel fold so far only seen in PRRSV and other nidoviruses. Cryo-electron tomographic studies have shown the three-dimensional organization of the PRRSV virion and suggest that the viral nucleocapsid has an asymmetric, linear arrangement, rather than the isometric core previously described. Together, these studies have revealed a closer structural relationship between arteri- and coronaviruses than previously anticipated.

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Section: Introductionmentioning

confidence: 99%

The structural biology of PRRSV

2010

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“…One such area is the region of the ORF1 replicase protein known as non-structural protein 2 (nsp2) (Snijder et al, 1995b;Tian et al, 2007;van Hemert and Snijder, 2008), the most variable section of the genome (Han et al, 2006). Based on considerable research on equine arteritis virus (EAV), the nsp2 protein is processed from the ORF1 protein by the action of the papain-like protease in nsp1␤ and the self-encoded PL2 cysteine protease, and acts as a co-factor for the nsp4 serine protease (3CL) in processing downstream viral cleavages (den Boon et al, 1995;Snijder et al, 1994Snijder et al, , 1995avan Hemert and Snijder, 2008;Wassenaar et al, 1997). In addition, EAV nsp2 induces double membrane vesicles, probably through its predicted transmembrane spanning regions, and serves as an anchor for the virus replication complex (Pedersen et al, 1999;Snijder et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

In vivo growth of porcine reproductive and respiratory syndrome virus engineered nsp2 deletion mutants

et al. 2010

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Prior studies on PRRSV strain VR-2332 non-structural protein 2 (nsp2) had shown that as much as 403 amino acids could be removed from the hypervariable region without losing virus viability in vitro. We utilized selected nsp2 deletion mutants to examine in vivo growth. Young swine (4 pigs/group; 5 control swine) were inoculated intramuscularly with one of 4 nsp2 deletion mutants (rΔ727-813, rΔ543-726, rΔ324-523, rΔ324-726) or full-length recombinant virus (rVR-2332). Serum samples were collected on various days post-inoculation and analyzed by HerdChek* ELISA, PRRSV real time RT-PCR, gamma interferon (IFN-γ) ELISA, and nucleotide sequence analysis of the entire nsp2 coding region. Tracheobronchial lymph node weight compared to body weight was recorded for each animal and used as a clinical measurement of viral pathogenesis. Results showed that all deletion mutants grew less robustly than full-length recombinant virus, yet all but the large deletion virus (rΔ324-726) recovered to parental viral RNA levels by study end. Swine receiving the rΔ727-813 mutants had a significant decrease in lymph node enlargement compared to rVR-2332. While swine infection with rVR-2332 caused a rapid rise in serum IFN-γ levels, the IFN-γ protein produced by infection with 3 of the 4 deletion mutant viruses was significantly reduced, perhaps due to differences in viral growth kinetics. The rΔ543-726 nsp2 mutant virus, although growth impaired, mimicked rVR-2332 in inducing a host serum IFN-γ response but exhibited a 2-week delay. Targeted sequencing showed that all deletions were stable in the region coding for nsp2 after one swine passage. The data suggested that the selected nsp2 deletion mutants were growth attenuated in swine, altered the induction of serum IFN-γ, an innate cytokine of unknown function in PRRSV clearance, and pointed to a domain that may influence tracheobronchial lymph node size.

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“…This activity cleaves off the nsp1a subunit and releases it from pp1a (Fig. 2) (Den Boon et al, 1995). The cleavage site by PLP1a is mapped to M180/A181 and confirmed by two independent studies using peptide sequencing and X-ray crystallographic study Sun et al, 2009).…”

Section: Nsp1amentioning

confidence: 61%

“…The PLP1b activity is responsible for the cleavage of nsp1b from nsp2 (Fig. 2) (Den Boon et al, 1995;Snijder et al, 1992;Ziebuhr et al, 2000). Thus, the release of nsp1b is dependent on the activities of PLP1a and PLP1b.…”

Section: Nsp1bmentioning

confidence: 99%

The viral innate immune antagonism and an alternative vaccine design for PRRS virus

Yoo

2017

Veterinary Microbiology

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Porcine reproductive and respiratory syndrome (PRRS) remains one of the most economically significant diseases in the swine industry worldwide. The current vaccines are less satisfactory to confer protections from heterologous infections and long-term persistence, and the need for better vaccines are urgent. The immunological hallmarks in PRRSV-infected pigs include the unusually poor production of type I interferons (IFNs-α/β) and the aberrant and delayed adaptive immune responses, indicating that PRRSV has the ability to suppress both innate and adaptive immune responses in the host. Type I IFNs are the potent antiviral cytokines and recent studies reveal their pleiotropic functions in the priming of expansion and maturation of adaptive immunity. Thus, IFN antagonism-negative PRRSV is hypothesized to be attenuated and to build effective and broad- spectrum innate and adaptive immune responses in pigs. Such vaccines are promising alternatives to traditional vaccines for PRRSV.

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Processing and evolution of the N-terminal region of the arterivirus replicase ORF1a protein: identification of two papainlike cysteine proteases

Cited by 172 publications

References 30 publications

The structural biology of PRRSV

The structural biology of PRRSV

In vivo growth of porcine reproductive and respiratory syndrome virus engineered nsp2 deletion mutants

The viral innate immune antagonism and an alternative vaccine design for PRRS virus

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