FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Journal of virology2008; 82(9); 4480-4491; doi: 10.1128/JVI.02756-07

Formation of the arterivirus replication/transcription complex: a key role for nonstructural protein 3 in the remodeling of intracellular membranes.

Abstract: The replication/transcription complex of the arterivirus equine arteritis virus (EAV) is associated with paired membranes and/or double-membrane vesicles (DMVs) that are thought to originate from the endoplasmic reticulum. Previously, coexpression of two putative transmembrane nonstructural proteins (nsp2 and nsp3) was found to suffice to induce these remarkable membrane structures, which are typical of arterivirus infection. Here, site-directed mutagenesis was used to investigate the role of nsp3 in more detail. Liberation of the hydrophobic N terminus of nsp3, which is normally achieved by cleavage of the nsp2/3 junction by the nsp2 protease, was nonessential for the formation of DMVs. However, the substitution of each of a cluster of four conserved cysteine residues, residing in a predicted luminal loop of nsp3, completely blocked DMV formation. Some of these mutant nsp3 proteins were also found to be highly cytotoxic, in particular, exerting a dramatic effect on the endoplasmic reticulum. The functionality of an engineered N glycosylation site in the cysteine-containing loop confirmed both its presence in the lumen and the transmembrane nature of nsp3. This mutant displayed an interesting intermediate phenotype in terms of DMV formation, with paired and curved membranes being formed, but DMV formation apparently being impaired. The effect of nsp3 mutations on replicase polyprotein processing was investigated, and several mutations were found to influence processing of the region downstream of nsp3 by the nsp4 main protease. When tested in an EAV reverse genetics system, none of the nsp3 mutations was tolerated, again underlining the crucial role of the protein in the arterivirus life cycle.
Get Full Text
Publication Date: 2008-02-27 PubMed ID: 18305048PubMed Central: PMC2293027DOI: 10.1128/JVI.02756-07Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on the Arterivirus Equine Arteritis Virus (EAV) and emphasizes the critical role of the nonstructural protein 3 (nsp3) in the formation of the virus’s replication/transcription complex, which is crucial for the virus’s life cycle. The study uses site-directed mutagenesis to investigate nsp3’s role in detail.

Arterivirus Equine Arteritis Virus (EAV)

  • This virus has a replication/transcription complex associated with paired membranes or double-membrane vesicles (DMVs) believed to originate from the endoplasmic reticulum, an organelle found in eukaryotic cells.
  • Previous studies had shown that coexpression of two transmembrane nonstructural proteins (nsp2 and nsp3) was enough to induce these DMVs, which are typical of arterivirus infection.

Role of Nonstructural Protein 3 (nsp3)

  • In this study, the researchers used site-directed mutagenesis, a method for making specific and intentional changes to the DNA sequence of a gene, to study the role of nsp3 in more detail.
  • The release of the hydrophobic N terminus of nsp3, usually achieved by the cleavage of the nsp2/3 junction by the nsp2 protease, was found to be nonessential for the formation of DMVs.
  • However, the substitution of each of four conserved cysteine residues found in a predicted luminal loop of nsp3 blocked DMV formation. Some of these mutant nsp3 proteins were found to be highly cytotoxic and affected the endoplasmic reticulum significantly.
  • The researchers verified the presence of the cysteine-containing loop in the lumen and confirmed the transmembrane nature of nsp3 by engineering an N glycosylation site in the loop. This mutant displayed an interesting intermediate phenotype with respect to DMV formation. Curved and paired membranes were formed, but DMV formation was impaired.

Importance in the Viral Life Cycle

  • The researchers also studied the effect of nsp3 mutations on the processing of the replicase polyprotein, a sequence of linked proteins required for virus replication. Several mutations were found to influence the processing of the region downstream of nsp3 by the nsp4 main protease.
  • The researchers tested these nsp3 mutations in an EAV reverse genetics system. None of these mutations were tolerated, reinforcing the importance of nsp3 in the virus’s life cycle.

Conclusion

  • This research provides key insights into the character of the arterivirus equine arteritis virus and the critical role of its nonstructural protein 3 in the virus’s formation and life cycle. This may inform future research and treatments for infections related to this virus.

Cite This Article

APA
Posthuma CC, Pedersen KW, Lu Z, Joosten RG, Roos N, Zevenhoven-Dobbe JC, Snijder EJ. (2008). Formation of the arterivirus replication/transcription complex: a key role for nonstructural protein 3 in the remodeling of intracellular membranes. J Virol, 82(9), 4480-4491. https://doi.org/10.1128/JVI.02756-07

Publication

Journal of virology
ISSN: 1098-5514
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 82
Issue: 9
Pages: 4480-4491

Researcher Affiliations

Posthuma, Clara C
  • Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, LUMC P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
Pedersen, Ketil W
    Lu, Zhengchun
      Joosten, Ruth G
        Roos, Norbert
          Zevenhoven-Dobbe, Jessika C
            Snijder, Eric J

              MeSH Terms

              • Animals
              • Arterivirus / chemistry
              • Arterivirus / physiology
              • Arterivirus / ultrastructure
              • Horses
              • Intracellular Membranes / virology
              • Multiprotein Complexes
              • Mutagenesis, Site-Directed
              • Transcription, Genetic
              • Viral Nonstructural Proteins / genetics
              • Viral Nonstructural Proteins / physiology
              • Virus Replication

              References

              This article includes 54 references
              1. Ahlquist P. Parallels among positive-strand RNA viruses, reverse-transcribing viruses and double-stranded RNA viruses.. Nat. Rev. Microbiol. 4371-382.
                pmc: PMC7097367pubmed: 16582931
              2. Ahlquist P, Noueiry AO, Lee WM, Kushner DB, Dye BT. Host factors in positive-strand RNA virus genome replication.. J. Virol. 778181-8186.
                pmc: PMC165243pubmed: 12857886
              3. Beerens N, Snijder EJ. An RNA pseudoknot in the 3′ end of the arterivirus genome has a critical role in regulating viral RNA synthesis.. J. Virol. 819426-9436.
                pmc: PMC1951461pubmed: 17581985
              4. Belov GA, tan-Bonnet N, Kovtunovych G, Jackson CL, Lippincott-Schwartz J, Ehrenfeld E. Hijacking components of the cellular secretory pathway for replication of poliovirus RNA.. J. Virol. 81558-567.
                pmc: PMC1797456pubmed: 17079330
              5. Bienz K, Egger D, Pfister T, Troxler M. Structural and functional characterization of the poliovirus replication complex.. J. Virol. 662740-2747.
                pmc: PMC241029pubmed: 1313898
              6. Bost AG, Prentice E, Denison MR. Mouse hepatitis virus replicase protein complexes are translocated to sites of M protein accumulation in the ERGIC at late times of infection.. Virology 28521-29.
                pmc: PMC7130751pubmed: 11414802
              7. Bredenbeek PJ, Frolov I, Rice CM, Schlesinger S. Sindbis virus expression vectors: packaging of RNA replicons by using defective helper RNAs.. J. Virol. 676439-6446.
                pmc: PMC238079pubmed: 8411346
              8. Field J, Nikawa J, Broek D, MacDonald B, Rodgers L, Wilson IA, Lerner RA, Wigler M. Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method.. Mol. Cell. Biol. 82159-2165.
                pmc: PMC363397pubmed: 2455217
              9. Frias-Staheli N, Giannakopoulos NV, Kikkert M, Taylor SL, Bridgen A, Paragas J, Richt JA, Rowland RR, Schmaljohn CS, Lenschow DJ, Snijder EJ, García-Sastre A, Virgin HW. Ovarian tumor domain-containing viral proteases evade ubiquitin- and ISG15-dependent innate immune responses.. Cell Host Microbe 2404-416.
                pmc: PMC2184509pubmed: 18078692
              10. Fuerst TR, Niles EG, Studier FW, Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase.. Proc. Natl. Acad. Sci. USA 838122-8126.
                pmc: PMC386879pubmed: 3095828
              11. Goldsmith CS, Tatti KM, Ksiazek TG, Rollin PE, Comer JA, Lee WW, Rota PA, Bankamp B, Bellini WJ, Zaki SR. Ultrastructural characterization of SARS coronavirus.. Emerg. Infect. Dis. 10320-326.
                pmc: PMC3322934pubmed: 15030705
              12. Gorbalenya AE, Enjuanes L, Ziebuhr J, Snijder EJ. Nidovirales: evolving the largest RNA virus genome.. Virus Res. 11717-37.
                pmc: PMC7114179pubmed: 16503362
              13. Gosert R, Kanjanahaluethai A, Egger D, Bienz K, Baker SC. RNA replication of mouse hepatitis virus takes place at double-membrane vesicles.. J. Virol. 763697-3708.
                pmc: PMC136101pubmed: 11907209
              14. Griffiths G, Simons K, Warren G, Tokuyasu KT. Immunoelectron microscopy using thin, frozen sections: application to studies of the intracellular transport of Semliki Forest virus spike glycoproteins.. Methods Enzymol. 96466-485.
                pubmed: 6656640
              15. Harcourt BH, Jukneliene D, Kanjanahaluethai A, Bechill J, Severson KM, Smith CM, Rota PA, Baker SC. Identification of severe acute respiratory syndrome coronavirus replicase products and characterization of papain-like protease activity.. J. Virol. 7813600-13612.
                pmc: PMC533933pubmed: 15564471
              16. Ivanov KA, Thiel V, Dobbe JC, van der Meer Y, Snijder EJ, Ziebuhr J. Multiple enzymatic activities associated with severe acute respiratory syndrome coronavirus helicase.. J. Virol. 785619-5632.
                pmc: PMC415832pubmed: 15140959
              17. Jang SK, Krausslich HG, Nicklin MJ, Duke GM, Palmenberg AC, Wimmer E. A segment of the 5′ nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation.. J. Virol. 622636-2643.
                pmc: PMC253694pubmed: 2839690
              18. Kanjanahaluethai A, Chen Z, Jukneliene D, Baker SC. Membrane topology of murine coronavirus replicase nonstructural protein 3.. Virology 361391-401.
                pmc: PMC1925034pubmed: 17222884
              19. Kirkegaard K, Taylor MP, Jackson WT. Cellular autophagy: surrender, avoidance and subversion by microorganisms.. Nat. Rev. Microbiol. 2301-314.
                pmc: PMC7097095pubmed: 15031729
              20. Kopek BG, Perkins G, Miller DJ, Ellisman MH, Ahlquist P. Three-dimensional analysis of a viral RNA replication complex reveals a virus-induced mini-organelle.. PLoS Biol. 5e220.
                pmc: PMC1945040pubmed: 17696647
              21. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227680-685.
                pubmed: 5432063
              22. Mackenzie J. Wrapping things up about virus RNA replication.. Traffic 6967-977.
                pmc: PMC7169867pubmed: 16190978
              23. Makarova KS, Aravind L, Koonin EV. A novel superfamily of predicted cysteine proteases from eukaryotes, viruses and Chlamydia pneumoniae.. Trends Biochem. Sci. 2550-52.
                pubmed: 10664582
              24. Netherton C, Moffat K, Brooks E, Wileman T. A guide to viral inclusions, membrane rearrangements, factories, and viroplasm produced during virus replication.. Adv. Virus Res. 70101-182.
                pmc: PMC7112299pubmed: 17765705
              25. Novoa RR, Calderita G, Arranz R, Fontana J, Granzow H, Risco C. Virus factories: associations of cell organelles for viral replication and morphogenesis.. Biol. Cell 97147-172.
                pmc: PMC7161905pubmed: 15656780
              26. Oostra M, Te Lintelo EG, Deijs M, Verheije MH, Rottier PJ, de Haan CA. Localization and membrane topology of the coronavirus nonstructural protein 4: involvement of the early secretory pathway in replication.. J. Virol. 8112323-12336.
                pmc: PMC2168994pubmed: 17855519
              27. Pasternak AO, Spaan WJM, Snijder EJ. Nidovirus transcription: how to make sense…?. J. Gen. Virol. 871403-1421.
                pubmed: 16690906
              28. Pedersen KW, van der Meer Y, Roos N, Snijder EJ. Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex.. J. Virol. 732016-2026.
                pmc: PMC104444pubmed: 9971782
              29. Prentice E, Jerome WG, Yoshimori T, Mizushima N, Denison MR. Coronavirus replication complex formation utilizes components of cellular autophagy.. J. Biol. Chem. 27910136-10141.
                pmc: PMC7957857pubmed: 14699140
              30. Roosendaal J, Westaway EG, Khromykh A, Mackenzie JM. Regulated cleavages at the West Nile virus NS4A-2K-NS4B junctions play a major role in rearranging cytoplasmic membranes and Golgi trafficking of the NS4A protein.. J. Virol. 804623-4632.
                pmc: PMC1472005pubmed: 16611922
              31. Salonen A, Ahola T, Kaariainen L. Viral RNA replication in association with cellular membranes.. Curr. Top. Microbiol. Immunol. 285139-173.
                pmc: PMC7120253pubmed: 15609503
              32. Sawicki SG, Sawicki DL, Siddell SG. A contemporary view of coronavirus transcription.. J. Virol. 8120-29.
                pmc: PMC1797243pubmed: 16928755
              33. Shi ST, Schiller JJ, Kanjanahaluethai A, Baker SC, Oh JW, Lai MM. Colocalization and membrane association of murine hepatitis virus gene 1 products and de novo-synthesized viral RNA in infected cells.. J. Virol. 735957-5969.
                pmc: PMC112657pubmed: 10364348
              34. Snijder EJ, Bredenbeek PJ, Dobbe JC, Thiel V, Ziebuhr J, Poon LLM, Guan Y, Rozanov M, Spaan WJM, Gorbalenya AE. Unique and conserved features of genome and proteome of SARS-coronavirus, an early split-off from the coronavirus group 2 lineage.. J. Mol. Biol. 331991-1004.
                pmc: PMC7159028pubmed: 12927536
              35. Snijder EJ, Siddell SG, Gorbalenya AE. The order Nidovirales, p. 390-404. In B. W. Mahy and V. ter Meulen (ed.), Topley and Wilson's microbiology and microbial infections: virology.. .
              36. Snijder EJ, van der Meer Y, Zevenhoven-Dobbe JC, Onderwater JJM, van der Meulen J, Koerten HK, Mommaas AM. Ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex.. J. Virol. 805927-5940.
                pmc: PMC1472606pubmed: 16731931
              37. Snijder EJ, van Tol H, Roos N, Pedersen KW. Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex.. J. Gen. Virol. 82985-994.
                pubmed: 11297673
              38. Snijder EJ, Wassenaar ALM, Spaan WJM. Proteolytic processing of the replicase ORF1a protein of equine arteritis virus.. J. Virol. 685755-5764.
                pmc: PMC236979pubmed: 8057457
              39. Snijder EJ, Wassenaar ALM, Spaan WJM, Gorbalenya AE. The arterivirus nsp2 protease. An unusual cysteine protease with primary structure similarities to both papain-like and chymotrypsin-like proteases.. J. Biol. Chem. 27016671-16676.
                pubmed: 7622476
              40. Snijder EJ, Wassenaar ALM, van Dinten LC, Spaan WJM, Gorbalenya AE. The arterivirus nsp4 protease is the prototype of a novel group of chymotrypsin-like enzymes, the 3C-like serine proteases.. J. Biol. Chem. 2714864-4871.
                pubmed: 8617757
              41. Sparks JS, Lu X, Denison MR. Genetic analysis of murine hepatitis virus nsp4 in virus replication.. J. Virol. 8112554-12563.
                pmc: PMC2169011pubmed: 17855548
              42. Stertz S, Reichelt M, Spiegel M, Kuri T, Martínez-Sobrido L, García-Sastre A, Weber F, Kochs G. The intracellular sites of early replication and budding of SARS-coronavirus.. Virology 361304-315.
                pmc: PMC7103305pubmed: 17210170
              43. Sulea T, Lindner HA, Menard R. Structural aspects of recently discovered viral deubiquitinating activities.. Biol. Chem. 387853-862.
                pubmed: 16913834
              44. van Aken D, Zevenhoven-Dobbe JC, Gorbalenya AE, Snijder EJ. Proteolytic maturation of replicase polyprotein pp1a by the nsp4 main proteinase is essential for equine arteritis virus replication and includes internal cleavage of nsp7.. J. Gen. Virol. 873473-3482.
                pubmed: 17098961
              45. van der Meer Y, Snijder EJ, Dobbe JC, Schleich S, Denison MR, Spaan WJM, Krijnse Locker J. Localization of mouse hepatitis virus nonstructural proteins and RNA synthesis indicates a role for late endosomes in viral replication.. J. Virol. 737641-7657.
                pmc: PMC104292pubmed: 10438855
              46. van der Meer Y, van Tol H, Krijnse Locker J, Snijder EJ. ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex.. J. Virol. 726689-6698.
                pmc: PMC109868pubmed: 9658116
              47. van Dinten LC, den Boon JA, Wassenaar ALM, Spaan WJM, Snijder EJ. An infectious arterivirus cDNA clone: identification of a replicase point mutation which abolishes discontinuous mRNA transcription.. Proc. Natl. Acad. Sci. USA 94991-996.
                pmc: PMC19627pubmed: 9023370
              48. van Dinten LC, Wassenaar ALM, Gorbalenya AE, Spaan WJM, Snijder EJ. Processing of the equine arteritis virus replicase ORF1b protein: identification of cleavage products containing the putative viral polymerase and helicase domains.. J. Virol. 706625-6633.
                pmc: PMC190703pubmed: 8794297
              49. van Marle G, van Dinten LC, Luytjes W, Spaan WJM, Snijder EJ. Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis.. J. Virol. 735274-5281.
                pmc: PMC112582pubmed: 10364273
              50. Vaux D, Tooze J, Fuller S. Identification by anti-idiotypic antibodies of an intracellular membrane protein that recognizes a mammalian endoplasmic reticulum retention signal.. Nature 345495-502.
                pubmed: 2161500
              51. Wassenaar ALM, Spaan WJM, Gorbalenya AE, Snijder EJ. Alternative proteolytic processing of the arterivirus replicase ORF1a polyprotein: evidence that NSP2 acts as a cofactor for the NSP4 serine protease.. J. Virol. 719313-9322.
                pmc: PMC230234pubmed: 9371590
              52. Wessels E, Duijsings D, Niu TK, Neumann S, Oorschot VM, de Lange F, Lanke KH, Klumperman J, Henke A, Jackson CL, Melchers WJ, van Kuppeveld FJ. A viral protein that blocks Arf1-mediated COP-I assembly by inhibiting the guanine nucleotide exchange factor GBF1.. Dev. Cell 11191-201.
                pubmed: 16890159
              53. Wileman T. Aggresomes and autophagy generate sites for virus replication.. Science 312875-878.
                pubmed: 16690857
              54. Ziebuhr J, Snijder EJ, Gorbalenya AE. Virus-encoded proteinases and proteolytic processing in the Nidovirales.. J. Gen. Virol. 81853-879.
                pubmed: 10725411

              Citations

              This article has been cited 57 times.
              1. Soares R, Vieira CP, Vieira J. Patterns of Recombination in Coronaviruses. Int J Mol Sci 2025 Jun 11;26(12).
                doi: 10.3390/ijms26125595pubmed: 40565057google scholar: lookup
              2. Lavender AA, Solanke O, Tang H-Y, Peck HE, Vanover D, Santangelo PJ, Brinton MA. The SHFV nsp2 and nucleocapsid proteins recruit G3BP1 to sites of viral replication, but stress granules are not induced by the infection. J Virol 2025 Jul 22;99(7):e0079425.
                doi: 10.1128/jvi.00794-25pubmed: 40548742google scholar: lookup
              3. Wang X, Chen Y, Qi C, Li F, Zhang Y, Zhou J, Wu H, Zhang T, Qi A, Ouyang H, Xie Z, Pang D. Mechanism, structural and functional insights into nidovirus-induced double-membrane vesicles. Front Immunol 2024;15:1340332.
                doi: 10.3389/fimmu.2024.1340332pubmed: 38919631google scholar: lookup
              4. Babot M, Boulard Y, Agouda S, Pieri L, Fieulaine S, Bressanelli S, Gervais V. Oligomeric assembly of the C-terminal and transmembrane region of SARS-CoV-2 nsp3. J Virol 2024 Apr 16;98(4):e0157523.
                doi: 10.1128/jvi.01575-23pubmed: 38483167google scholar: lookup
              5. Liao Y, Wang H, Liao H, Sun Y, Tan L, Song C, Qiu X, Ding C. Classification, replication, and transcription of Nidovirales. Front Microbiol 2023;14:1291761.
                doi: 10.3389/fmicb.2023.1291761pubmed: 38328580google scholar: lookup
              6. Liu B, Luo L, Shi Z, Ju H, Yu L, Li G, Cui J. Research Progress of Porcine Reproductive and Respiratory Syndrome Virus NSP2 Protein. Viruses 2023 Nov 24;15(12).
                doi: 10.3390/v15122310pubmed: 38140551google scholar: lookup
              7. Zhou Z, Qiu Y, Ge X. The taxonomy, host range and pathogenicity of coronaviruses and other viruses in the Nidovirales order. Anim Dis 2021;1(1):5.
                doi: 10.1186/s44149-021-00005-9pubmed: 34778878google scholar: lookup
              8. Wang TY, Sun MX, Zhang HL, Wang G, Zhan G, Tian ZJ, Cai XH, Su C, Tang YD. Evasion of Antiviral Innate Immunity by Porcine Reproductive and Respiratory Syndrome Virus. Front Microbiol 2021;12:693799.
                doi: 10.3389/fmicb.2021.693799pubmed: 34512570google scholar: lookup
              9. Hu Y, Ke P, Gao P, Zhang Y, Zhou L, Ge X, Guo X, Han J, Yang H. Identification of an Intramolecular Switch That Controls the Interaction of Helicase nsp10 with Membrane-Associated nsp12 of Porcine Reproductive and Respiratory Syndrome Virus. J Virol 2021 Aug 10;95(17):e0051821.
                doi: 10.1128/JVI.00518-21pubmed: 34076477google scholar: lookup
              10. Wolff G, Melia CE, Snijder EJ, Bárcena M. Double-Membrane Vesicles as Platforms for Viral Replication. Trends Microbiol 2020 Dec;28(12):1022-1033.
                doi: 10.1016/j.tim.2020.05.009pubmed: 32536523google scholar: lookup
              11. Shang P, Yuan F, Misra S, Li Y, Fang Y. Hyper-phosphorylation of nsp2-related proteins of porcine reproductive and respiratory syndrome virus. Virology 2020 Apr;543:63-75.
                doi: 10.1016/j.virol.2020.01.018pubmed: 32174300google scholar: lookup
              12. Wang TY, Fang QQ, Cong F, Liu YG, Wang HM, Zhang HL, Tian ZJ, Tang YD, Cai XH. The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis. Emerg Microbes Infect 2019;8(1):1501-1510.
                doi: 10.1080/22221751.2019.1679010pubmed: 31631782google scholar: lookup
              13. Song J, Gao P, Kong C, Zhou L, Ge X, Guo X, Han J, Yang H. The nsp2 Hypervariable Region of Porcine Reproductive and Respiratory Syndrome Virus Strain JXwn06 Is Associated with Viral Cellular Tropism to Primary Porcine Alveolar Macrophages. J Virol 2019 Dec 15;93(24).
                doi: 10.1128/JVI.01436-19pubmed: 31554681google scholar: lookup
              14. Song J, Liu Y, Gao P, Hu Y, Chai Y, Zhou S, Kong C, Zhou L, Ge X, Guo X, Han J, Yang H. Mapping the Nonstructural Protein Interaction Network of Porcine Reproductive and Respiratory Syndrome Virus. J Virol 2018 Dec 15;92(24).
                doi: 10.1128/JVI.01112-18pubmed: 30282705google scholar: lookup
              15. Doyle N, Neuman BW, Simpson J, Hawes PC, Mantell J, Verkade P, Alrashedi H, Maier HJ. Infectious Bronchitis Virus Nonstructural Protein 4 Alone Induces Membrane Pairing. Viruses 2018 Sep 6;10(9).
                doi: 10.3390/v10090477pubmed: 30200673google scholar: lookup
              16. Nan H, Lan J, Tian M, Dong S, Tian J, Liu L, Xu X, Chen H. The Network of Interactions Among Porcine Reproductive and Respiratory Syndrome Virus Non-structural Proteins. Front Microbiol 2018;9:970.
                doi: 10.3389/fmicb.2018.00970pubmed: 29867873google scholar: lookup
              17. Rappe JCF, de Wilde A, Di H, Müller C, Stalder H, V'kovski P, Snijder E, Brinton MA, Ziebuhr J, Ruggli N, Thiel V. Antiviral activity of K22 against members of the order Nidovirales. Virus Res 2018 Feb 15;246:28-34.
                doi: 10.1016/j.virusres.2018.01.002pubmed: 29337162google scholar: lookup
              18. Oudshoorn D, Rijs K, Limpens RWAL, Groen K, Koster AJ, Snijder EJ, Kikkert M, Bárcena M. Expression and Cleavage of Middle East Respiratory Syndrome Coronavirus nsp3-4 Polyprotein Induce the Formation of Double-Membrane Vesicles That Mimic Those Associated with Coronaviral RNA Replication. mBio 2017 Nov 21;8(6).
                doi: 10.1128/mBio.01658-17pubmed: 29162711google scholar: lookup
              19. Tuijtel MW, Mulder AA, Posthuma CC, van der Hoeven B, Koster AJ, Bárcena M, Faas FGA, Sharp TH. Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision. Sci Rep 2017 Sep 5;7(1):10442.
                doi: 10.1038/s41598-017-10905-xpubmed: 28874723google scholar: lookup
              20. Shulla A, Randall G. (+) RNA virus replication compartments: a safe home for (most) viral replication. Curr Opin Microbiol 2016 Aug;32:82-88.
                doi: 10.1016/j.mib.2016.05.003pubmed: 27253151google scholar: lookup
              21. van der Hoeven B, Oudshoorn D, Koster AJ, Snijder EJ, Kikkert M, Bárcena M. Biogenesis and architecture of arterivirus replication organelles. Virus Res 2016 Jul 15;220:70-90.
                doi: 10.1016/j.virusres.2016.04.001pubmed: 27071852google scholar: lookup
              22. Romero-Brey I, Berger C, Kallis S, Kolovou A, Paul D, Lohmann V, Bartenschlager R. NS5A Domain 1 and Polyprotein Cleavage Kinetics Are Critical for Induction of Double-Membrane Vesicles Associated with Hepatitis C Virus Replication. mBio 2015 Jul 7;6(4):e00759.
                doi: 10.1128/mBio.00759-15pubmed: 26152585google scholar: lookup
              23. V'kovski P, Al-Mulla H, Thiel V, Neuman BW. New insights on the role of paired membrane structures in coronavirus replication. Virus Res 2015 Apr 16;202:33-40.
                doi: 10.1016/j.virusres.2014.12.021pubmed: 25550072google scholar: lookup
              24. Steinbach F, Westcott DG, McGowan SL, Grierson SS, Frossard JP, Choudhury B. Re-emergence of a genetic outlier strain of equine arteritis virus: Impact on phylogeny. Virus Res 2015 Apr 16;202:144-50.
                doi: 10.1016/j.virusres.2014.12.009pubmed: 25527462google scholar: lookup
              25. Maier HJ, Hawes PC, Keep SM, Britton P. Spherules and IBV. Bioengineered 2014 Sep-Oct;5(5):288-92.
                doi: 10.4161/bioe.29323pubmed: 25482229google scholar: lookup
              26. Gouttenoire J, Montserret R, Paul D, Castillo R, Meister S, Bartenschlager R, Penin F, Moradpour D. Aminoterminal amphipathic α-helix AH1 of hepatitis C virus nonstructural protein 4B possesses a dual role in RNA replication and virus production. PLoS Pathog 2014 Oct;10(10):e1004501.
                doi: 10.1371/journal.ppat.1004501pubmed: 25392992google scholar: lookup
              27. Romero-Brey I, Bartenschlager R. Membranous replication factories induced by plus-strand RNA viruses. Viruses 2014 Jul 22;6(7):2826-57.
                doi: 10.3390/v6072826pubmed: 25054883google scholar: lookup
              28. Neuman BW, Angelini MM, Buchmeier MJ. Does form meet function in the coronavirus replicative organelle?. Trends Microbiol 2014 Nov;22(11):642-7.
                doi: 10.1016/j.tim.2014.06.003pubmed: 25037114google scholar: lookup
              29. Wang L, Zhou L, Zhang H, Li Y, Ge X, Guo X, Yu K, Yang H. Interactome profile of the host cellular proteins and the nonstructural protein 2 of porcine reproductive and respiratory syndrome virus. PLoS One 2014;9(6):e99176.
                doi: 10.1371/journal.pone.0099176pubmed: 24901321google scholar: lookup
              30. Lundin A, Dijkman R, Bergström T, Kann N, Adamiak B, Hannoun C, Kindler E, Jónsdóttir HR, Muth D, Kint J, Forlenza M, Müller MA, Drosten C, Thiel V, Trybala E. Targeting membrane-bound viral RNA synthesis reveals potent inhibition of diverse coronaviruses including the middle East respiratory syndrome virus. PLoS Pathog 2014 May;10(5):e1004166.
                doi: 10.1371/journal.ppat.1004166pubmed: 24874215google scholar: lookup
              31. Bailey-Elkin BA, van Kasteren PB, Snijder EJ, Kikkert M, Mark BL. Viral OTU deubiquitinases: a structural and functional comparison. PLoS Pathog 2014 Mar;10(3):e1003894.
                doi: 10.1371/journal.ppat.1003894pubmed: 24676359google scholar: lookup
              32. Angelini MM, Neuman BW, Buchmeier MJ. Untangling membrane rearrangement in the nidovirales. DNA Cell Biol 2014 Mar;33(3):122-7.
                doi: 10.1089/dna.2013.2304pubmed: 24410069google scholar: lookup
              33. Yun SI, Lee YM. Overview: Replication of porcine reproductive and respiratory syndrome virus. J Microbiol 2013 Dec;51(6):711-23.
                doi: 10.1007/s12275-013-3431-zpubmed: 24385346google scholar: lookup
              34. Paul D, Bartenschlager R. Architecture and biogenesis of plus-strand RNA virus replication factories. World J Virol 2013 May 12;2(2):32-48.
                doi: 10.5501/wjv.v2.i2.32pubmed: 24175228google scholar: lookup
              35. Angelini MM, Akhlaghpour M, Neuman BW, Buchmeier MJ. Severe acute respiratory syndrome coronavirus nonstructural proteins 3, 4, and 6 induce double-membrane vesicles. mBio 2013 Aug 13;4(4).
                doi: 10.1128/mBio.00524-13pubmed: 23943763google scholar: lookup
              36. Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
                doi: 10.1016/j.vetmic.2013.06.015pubmed: 23891306google scholar: lookup
              37. van Kasteren PB, Bailey-Elkin BA, James TW, Ninaber DK, Beugeling C, Khajehpour M, Snijder EJ, Mark BL, Kikkert M. Deubiquitinase function of arterivirus papain-like protease 2 suppresses the innate immune response in infected host cells. Proc Natl Acad Sci U S A 2013 Feb 26;110(9):E838-47.
                doi: 10.1073/pnas.1218464110pubmed: 23401522google scholar: lookup
              38. Zeng X, Carlin CR. Host cell autophagy modulates early stages of adenovirus infections in airway epithelial cells. J Virol 2013 Feb;87(4):2307-19.
                doi: 10.1128/JVI.02014-12pubmed: 23236070google scholar: lookup
              39. Maier HJ, Britton P. Involvement of autophagy in coronavirus replication. Viruses 2012 Nov 30;4(12):3440-51.
                doi: 10.3390/v4123440pubmed: 23202545google scholar: lookup
              40. Hagemeijer MC, Rottier PJ, de Haan CA. Biogenesis and dynamics of the coronavirus replicative structures. Viruses 2012 Nov 21;4(11):3245-69.
                doi: 10.3390/v4113245pubmed: 23202524google scholar: lookup
              41. Zhang J, Go YY, Huang CM, Meade BJ, Lu Z, Snijder EJ, Timoney PJ, Balasuriya UB. Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus. Clin Vaccine Immunol 2012 Aug;19(8):1312-21.
                doi: 10.1128/CVI.00302-12pubmed: 22739697google scholar: lookup
              42. Netherton CL, Wileman T. Virus factories, double membrane vesicles and viroplasm generated in animal cells. Curr Opin Virol 2011 Nov;1(5):381-7.
                doi: 10.1016/j.coviro.2011.09.008pubmed: 22440839google scholar: lookup
              43. Knoops K, Bárcena M, Limpens RW, Koster AJ, Mommaas AM, Snijder EJ. Ultrastructural characterization of arterivirus replication structures: reshaping the endoplasmic reticulum to accommodate viral RNA synthesis. J Virol 2012 Mar;86(5):2474-87.
                doi: 10.1128/JVI.06677-11pubmed: 22190716google scholar: lookup
              44. Dreux M, Chisari FV. Impact of the autophagy machinery on hepatitis C virus infection. Viruses 2011 Aug;3(8):1342-57.
                doi: 10.3390/v3081342pubmed: 21994783google scholar: lookup
              45. Stapleford KA, Miller DJ. Role of cellular lipids in positive-sense RNA virus replication complex assembly and function. Viruses 2010 May;2(5):1055-1068.
                doi: 10.3390/v2051055pubmed: 21994671google scholar: lookup
              46. Fang Y, Snijder EJ. The PRRSV replicase: exploring the multifunctionality of an intriguing set of nonstructural proteins. Virus Res 2010 Dec;154(1-2):61-76.
                doi: 10.1016/j.virusres.2010.07.030pubmed: 20696193google scholar: lookup
              47. Dokland T. The structural biology of PRRSV. Virus Res 2010 Dec;154(1-2):86-97.
                doi: 10.1016/j.virusres.2010.07.029pubmed: 20692304google scholar: lookup
              48. Nedialkova DD, Gorbalenya AE, Snijder EJ. Arterivirus Nsp1 modulates the accumulation of minus-strand templates to control the relative abundance of viral mRNAs. PLoS Pathog 2010 Feb 19;6(2):e1000772.
                doi: 10.1371/journal.ppat.1000772pubmed: 20174607google scholar: lookup
              49. Sir D, Ou JH. Autophagy in viral replication and pathogenesis. Mol Cells 2010 Jan;29(1):1-7.
                doi: 10.1007/s10059-010-0014-2pubmed: 20077024google scholar: lookup
              50. Beura LK, Sarkar SN, Kwon B, Subramaniam S, Jones C, Pattnaik AK, Osorio FA. Porcine reproductive and respiratory syndrome virus nonstructural protein 1beta modulates host innate immune response by antagonizing IRF3 activation. J Virol 2010 Feb;84(3):1574-84.
                doi: 10.1128/JVI.01326-09pubmed: 19923190google scholar: lookup
              51. Gadlage MJ, Sparks JS, Beachboard DC, Cox RG, Doyle JD, Stobart CC, Denison MR. Murine hepatitis virus nonstructural protein 4 regulates virus-induced membrane modifications and replication complex function. J Virol 2010 Jan;84(1):280-90.
                doi: 10.1128/JVI.01772-09pubmed: 19846526google scholar: lookup
              52. Mizui T, Yamashina S, Tanida I, Takei Y, Ueno T, Sakamoto N, Ikejima K, Kitamura T, Enomoto N, Sakai T, Kominami E, Watanabe S. Inhibition of hepatitis C virus replication by chloroquine targeting virus-associated autophagy. J Gastroenterol 2010 Feb;45(2):195-203.
                doi: 10.1007/s00535-009-0132-9pubmed: 19760134google scholar: lookup
              53. Denison MR. Seeking membranes: positive-strand RNA virus replication complexes. PLoS Biol 2008 Oct 28;6(10):e270.
                doi: 10.1371/journal.pbio.0060270pubmed: 18959488google scholar: lookup
              54. Wei T, Wang A. Biogenesis of cytoplasmic membranous vesicles for plant potyvirus replication occurs at endoplasmic reticulum exit sites in a COPI- and COPII-dependent manner. J Virol 2008 Dec;82(24):12252-64.
                doi: 10.1128/JVI.01329-08pubmed: 18842721google scholar: lookup
              55. Oostra M, Hagemeijer MC, van Gent M, Bekker CP, te Lintelo EG, Rottier PJ, de Haan CA. Topology and membrane anchoring of the coronavirus replication complex: not all hydrophobic domains of nsp3 and nsp6 are membrane spanning. J Virol 2008 Dec;82(24):12392-405.
                doi: 10.1128/JVI.01219-08pubmed: 18842706google scholar: lookup
              56. Knoops K, Kikkert M, Worm SH, Zevenhoven-Dobbe JC, van der Meer Y, Koster AJ, Mommaas AM, Snijder EJ. SARS-coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. PLoS Biol 2008 Sep 16;6(9):e226.
                doi: 10.1371/journal.pbio.0060226pubmed: 18798692google scholar: lookup
              57. You JH, Howell G, Pattnaik AK, Osorio FA, Hiscox JA. A model for the dynamic nuclear/nucleolar/cytoplasmic trafficking of the porcine reproductive and respiratory syndrome virus (PRRSV) nucleocapsid protein based on live cell imaging. Virology 2008 Aug 15;378(1):34-47.
                doi: 10.1016/j.virol.2008.04.037pubmed: 18550142google scholar: lookup
              Subscribe to our newsletter
              Join 99,780 horse owners like you who receive our email updates on horse health and nutrition.