The Porcine Deltacoronavirus Replication Organelle Comprises Double-Membrane Vesicles and Zippered Endoplasmic Reticulum with Double-Membrane Spherules
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Virus
2.2. Reverse Transcription and Quantitative Polymerase Chain Reaction
2.3. Western Blot
2.4. Virus Growth Curve and Titration by TCID50
2.5. Immunofluorescence
2.6. Transmission Electron Microscopy
3. Results
3.1. Kinetics of Porcine deltacoronavirus Replication in LLC-PK1 Cells
3.2. Visualizing Porcine deltacoronavirus RNA Synthesis
3.3. PDCoV ROs Include Double-Membrane Vesicles and Zippered ER with Double-Membrane Spherules
3.4. PDCoV ROs, Including Zippered ER and Double-Membrane Spherules, are Visible from 6 hpi
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Grimley, P.M.; Berezesky, I.K.; Friedman, R.M. Cytoplasmic structures associated with an arbovirus infection: Loci of viral ribonucleic acid synthesis. J. Virol. 1968, 2, 1326–1338. [Google Scholar] [PubMed]
- Grimley, P.M.; Levin, J.G.; Berezesky, I.K.; Friedman, R.M. Specific membranous structures associated with the replication of group A arboviruses. J. Virol. 1972, 10, 492–503. [Google Scholar] [PubMed]
- Kopek, B.G.; Perkins, G.; Miller, D.J.; Ellisman, M.H.; Ahlquist, P. Three-dimensional analysis of a viral RNA replication complex reveals a virus-induced mini-organelle. PLoS Biol. 2007, 5, e220. [Google Scholar] [CrossRef] [PubMed]
- Kujala, P.; Ikaheimonen, A.; Ehsani, N.; Vihinen, H.; Auvinen, P.; Kaariainen, L. Biogenesis of the Semliki Forest virus RNA replication complex. J. Virol. 2001, 75, 3873–3884. [Google Scholar] [CrossRef] [PubMed]
- Avila-Perez, G.; Rejas, M.T.; Rodriguez, D. Ultrastructural characterization of membranous torovirus replication factories. Cell Microbiol. 2016, 18, 1691–1708. [Google Scholar] [CrossRef] [Green Version]
- Belov, G.A.; Nair, V.; Hansen, B.T.; Hoyt, F.H.; Fischer, E.R.; Ehrenfeld, E. Complex dynamic development of poliovirus membranous replication complexes. J. Virol. 2012, 86, 302–312. [Google Scholar] [CrossRef] [PubMed]
- Gosert, R.; Egger, D.; Lohmann, V.; Bartenschlager, R.; Blum, H.E.; Bienz, K.; Moradpour, D. Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons. J. Virol. 2003, 77, 5487–5492. [Google Scholar] [CrossRef]
- Knoops, K.; Barcena, M.; Limpens, R.W.; Koster, A.J.; Mommaas, A.M.; Snijder, E.J. Ultrastructural characterization of arterivirus replication structures: Reshaping the endoplasmic reticulum to accommodate viral RNA synthesis. J. Virol. 2012, 86, 2474–2487. [Google Scholar] [CrossRef]
- Limpens, R.W.A.L.; van der Schaar, H.M.; Kumar, D.; Koster, A.J.; Snijder, E.J.; van Kuppeveld, F.J.M.; Bárcena, M. The transformation of enterovirus replication structures: A three-dimensional study of single- and double-membrane compartments. MBio 2011, 2, e00166-11. [Google Scholar] [CrossRef]
- Monaghan, P.; Cook, H.; Jackson, T.; Ryan, M.; Wileman, T. The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells. J. Gen. Virol. 2004, 85, 933–946. [Google Scholar] [CrossRef]
- Gillespie, L.K.; Hoenen, A.; Morgan, G.; Mackenzie, J.M. The Endoplasmic Reticulum Provides the Membrane Platform for Biogenesis of the Flavivirus Replication Complex. J. Virol. 2010, 84, 10438–10447. [Google Scholar] [CrossRef] [Green Version]
- Offerdahl, D.K.; Dorward, D.W.; Hansen, B.T.; Bloom, M.E. A Three-Dimensional Comparison of Tick-Borne Flavivirus Infection in Mammalian and Tick Cell Lines. PLoS ONE 2012, 7, e47912. [Google Scholar] [CrossRef] [PubMed]
- Schwartz, M.; Chen, J.; Lee, W.-M.; Janda, M.; Ahlquist, P. Alternate, virus-induced membrane rearrangements support positive-strand RNA virus genome replication. Proc. Natl. Acad. Sci. USA 2004, 101, 11263–11268. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Welsch, S.; Miller, S.; Romero-Brey, I.; Merz, A.; Bleck, C.K.; Walther, P.; Fuller, S.D.; Antony, C.; Krijnse-Locker, J.; Bartenschlager, R. Composition and three-dimensional architecture of the dengue virus replication and assembly sites. Cell Host Microbe 2009, 5, 365–375. [Google Scholar] [CrossRef] [PubMed]
- Westaway, E.G.; Mackenzie, J.M.; Kenney, M.T.; Jones, M.K.; Khromykh, A.A. Ultrastructure of Kunjin virus-infected cells: Colocalization of NS1 and NS3 with double-stranded RNA, and of NS2B with NS3, in virus-induced membrane structures. J. Virol. 1997, 71, 6650–6661. [Google Scholar] [PubMed]
- Ksiazek, T.G.; Erdman, D.; Goldsmith, C.S.; Zaki, S.R.; Peret, T.; Emery, S.; Tong, S.; Urbani, C.; Comer, J.A.; Lim, W.; et al. A novel coronavirus associated with severe acute respiratory syndrome. N. Engl. J. Med. 2003, 348, 1953–1966. [Google Scholar] [CrossRef]
- Zaki, A.M.; van Boheemen, S.; Bestebroer, T.M.; Osterhaus, A.D.; Fouchier, R.A. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N. Engl. J. Med. 2012, 367, 1814–1820. [Google Scholar] [CrossRef]
- Zhou, L.; Li, Q.N.; Su, J.N.; Chen, G.H.; Wu, Z.X.; Luo, Y.; Wu, R.T.; Sun, Y.; Lan, T.; Ma, J.Y. The re-emerging of SADS-CoV infection in pig herds in Southern China. Transbound Emerg. Dis. 2019, 6, 2180–2183. [Google Scholar] [CrossRef]
- Zhou, P.; Fan, H.; Lan, T.; Yang, X.-L.; Shi, W.-F.; Zhang, W.; Zhu, Y.; Zhang, Y.-W.; Xie, Q.-M.; Mani, S.; et al. Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin. Nature 2018, 556, 255–258. [Google Scholar] [CrossRef]
- Knoops, K.; Kikkert, M.; van den Worm, S.H.E.; Zevenhoven-Dobbe, J.C.; van der Meer, Y.; Koster, A.J.; Mommaas, A.M.; Snijder, E.J. SARS-Coronavirus Replication Is Supported by a Reticulovesicular Network of Modified Endoplasmic Reticulum. PLoS Biol. 2008, 6, e226. [Google Scholar] [CrossRef]
- Ulasli, M.; Verheije, M.H.; de Haan, C.A.M.; Reggiori, F. Qualitative and quantitative ultrastructural analysis of the membrane rearrangements induced by coronavirus. Cell. Microbiol. 2010, 12, 844–861. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhou, X.; Cong, Y.; Veenendaal, T.; Klumperman, J.; Shi, D.; Mari, M.; Reggiori, F. Ultrastructural Characterization of Membrane Rearrangements Induced by Porcine Epidemic Diarrhea Virus Infection. Viruses 2017, 9, 251. [Google Scholar] [CrossRef] [PubMed]
- de Wilde, A.H.; Raj, V.S.; Oudshoorn, D.; Bestebroer, T.M.; van Nieuwkoop, S.; Limpens, R.W.; Posthuma, C.C.; van der Meer, Y.; Barcena, M.; Haagmans, B.L.; et al. MERS-coronavirus replication induces severe in vitro cytopathology and is strongly inhibited by cyclosporin A or interferon-alpha treatment. J. Gen. Virol. 2013, 94, 1749–1760. [Google Scholar] [CrossRef] [PubMed]
- Maier, H.J.; Hawes, P.C.; Cottam, E.M.; Mantell, J.; Verkade, P.; Monaghan, P.; Wileman, T.; Britton, P. Infectious bronchitis virus generates spherules from zippered endoplasmic reticulum membranes. MBio 2013, 4, e00801-13. [Google Scholar] [CrossRef]
- Maier, H.J.; Neuman, B.W.; Bickerton, E.; Keep, S.M.; Alrashedi, H.; Hall, R.; Britton, P. Extensive coronavirus-induced membrane rearrangements are not a determinant of pathogenicity. Sci. Rep. 2016, 6, 27126. [Google Scholar] [CrossRef]
- Woo, P.C.; Lau, S.K.; Lam, C.S.; Lau, C.C.; Tsang, A.K.; Lau, J.H.; Bai, R.; Teng, J.L.; Tsang, C.C.; Wang, M.; et al. Discovery of seven novel Mammalian and avian coronaviruses in the genus deltacoronavirus supports bat coronaviruses as the gene source of alphacoronavirus and betacoronavirus and avian coronaviruses as the gene source of gammacoronavirus and deltacoronavirus. J. Virol. 2012, 86, 3995–4008. [Google Scholar]
- Dong, N.; Fang, L.; Zeng, S.; Sun, Q.; Chen, H.; Xiao, S. Porcine Deltacoronavirus in Mainland China. Emerg. Infect. Dis. 2015, 21, 2254–2255. [Google Scholar] [CrossRef]
- Lee, J.H.; Chung, H.C.; Nguyen, V.G.; Moon, H.J.; Kim, H.K.; Park, S.J.; Lee, C.H.; Lee, G.E.; Park, B.K. Detection and Phylogenetic Analysis of Porcine Deltacoronavirus in Korean Swine Farms, 2015. Transbound Emerg. Dis. 2016, 63, 248–252. [Google Scholar] [CrossRef]
- Lorsirigool, A.; Saeng-Chuto, K.; Temeeyasen, G.; Madapong, A.; Tripipat, T.; Wegner, M.; Tuntituvanont, A.; Intrakamhaeng, M.; Nilubol, D. The first detection and full-length genome sequence of porcine deltacoronavirus isolated in Lao PDR. Arch Virol. 2016, 161, 2909–2911. [Google Scholar] [CrossRef] [Green Version]
- Wang, L.; Byrum, B.; Zhang, Y. Detection and genetic characterization of deltacoronavirus in pigs, Ohio, USA, 2014. Emerg. Infect. Dis. 2014, 20, 1227–1230. [Google Scholar] [CrossRef]
- Wang, M.; Wang, Y.; Baloch, A.R.; Pan, Y.; Tian, L.; Xu, F.; Shivaramu, S.; Chen, S.; Zeng, Q. Detection and genetic characterization of porcine deltacoronavirus in Tibetan pigs surrounding the Qinghai-Tibet Plateau of China. Transbound Emerg. Dis. 2018, 65, 363–369. [Google Scholar] [CrossRef] [PubMed]
- Jung, K.; Hu, H.; Eyerly, B.; Lu, Z.; Chepngeno, J.; Saif, L.J. Pathogenicity of 2 porcine deltacoronavirus strains in gnotobiotic pigs. Emerg. Infect. Dis. 2015, 21, 650–654. [Google Scholar] [CrossRef] [PubMed]
- Dong, N.; Fang, L.; Yang, H.; Liu, H.; Du, T.; Fang, P.; Wang, D.; Chen, H.; Xiao, S. Isolation, genomic characterization, and pathogenicity of a Chinese porcine deltacoronavirus strain CHN-HN-2014. Vet. Microbiol. 2016, 196, 98–106. [Google Scholar] [CrossRef] [PubMed]
- Hu, H.; Jung, K.; Vlasova, A.N.; Chepngeno, J.; Lu, Z.; Wang, Q.; Saif, L.J. Isolation and characterization of porcine deltacoronavirus from pigs with diarrhea in the United States. J. Clin. Microbiol. 2015, 53, 1537–1548. [Google Scholar] [CrossRef] [PubMed]
- Jang, G.; Kim, S.H.; Lee, Y.J.; Kim, S.; Lee, D.S.; Lee, K.K.; Lee, C. Isolation and characterization of Korean porcine deltacoronavirus strain KNU16-07. J. Vet. Sci. 2018, 19, 577–581. [Google Scholar] [CrossRef]
- Qin, P.; Du, E.Z.; Luo, W.T.; Yang, Y.L.; Zhang, Y.Q.; Wang, B.; Huang, Y.W. Characteristics of the Life Cycle of Porcine Deltacoronavirus (PDCoV) In Vitro: Replication Kinetics, Cellular Ultrastructure and Virion Morphology, and Evidence of Inducing Autophagy. Viruses 2019, 11, 455. [Google Scholar] [CrossRef]
- Hull, R.N.; Cherry, W.R.; Weaver, G.W. The origin and characteristics of a pig kidney cell strain, LLC-PK. Vitro 1976, 12, 670–677. [Google Scholar] [CrossRef]
- Marthaler, D.; Jiang, Y.; Collins, J.; Rossow, K. Complete Genome Sequence of Strain SDCV/USA/Illinois121/2014, a Porcine Deltacoronavirus from the United States. Genome Announc. 2014, 2, e00218-14. [Google Scholar] [CrossRef] [Green Version]
- Hagemeijer, M.C.; Vonk, A.M.; Monastyrska, I.; Rottier, P.J.; de Haan, C.A. Visualizing coronavirus RNA synthesis in time by using click chemistry. J. Virol. 2012, 86, 5808–5816. [Google Scholar] [CrossRef]
- Rouiller, I.; Brookes, S.M.; Hyatt, A.D.; Windsor, M.; Wileman, T. African swine fever virus is wrapped by the endoplasmic reticulum. J. Virol. 1998, 72, 2373–2387. [Google Scholar]
- Sola, I.; Galán, C.; Mateos-Gómez, P.A.; Palacio, L.; Zúñiga, S.; Cruz, J.L.; Almazán, F.; Enjuanes, L. The polypyrimidine tract-binding protein affects coronavirus RNA accumulation levels and relocalizes viral RNAs to novel cytoplasmic domains different from replication-transcription sites. J. Virol. 2011, 85, 5136–5149. [Google Scholar] [CrossRef] [PubMed]
- Gui, M.; Liu, X.; Guo, D.Y.; Zhang, Z.; Yin, C.C.; Chen, Y.; Xiang, Y. Electron microscopy studies of the coronavirus ribonucleoprotein complex. Protein Cell 2017, 8, 219–224. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gosert, R.; Kanjanahaluethai, A.; Egger, D.; Bienz, K.; Baker, S.C. RNA Replication of Mouse Hepatitis Virus Takes Place at Double-Membrane Vesicles. J. Virol. 2002, 76, 3697–3708. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Snijder, E.J.; van der Meer, Y.; Zevenhoven-Dobbe, J.; Onderwater, J.J.M.; van der Meulen, J.; Koerten, H.K.; Mommaas, A.M. Ultrastructure and Origin of Membrane Vesicles Associated with the Severe Acute Respiratory Syndrome Coronavirus Replication Complex. J. Virol. 2006, 80, 5927–5940. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sawicki, S.G.; Sawicki, D.L. Coronavirus minus-strand RNA synthesis and effect of cycloheximide on coronavirus RNA synthesis. J. Virol. 1986, 57, 328–334. [Google Scholar] [PubMed]
- Sawicki, S.G.; Sawicki, D.L. Coronavirus transcription: A perspective. Curr. Top. Microbiol. Immunol. 2005, 287, 31–55. [Google Scholar]
- Sawicki, S.G.; Sawicki, D.L.; Siddell, S.G. A contemporary view of coronavirus transcription. J. Virol. 2007, 81, 20–29. [Google Scholar] [CrossRef]
- Goldsmith, C.S.; Tatti, K.M.; Ksiazek, T.G.; Rollin, P.E.; Comer, J.A.; Lee, W.W.; Rota, P.A.; Bankamp, B.; Bellini, W.J.; Zaki, S.R. Ultrastructural characterization of SARS coronavirus. Emerg. Infect. Dis. 2004, 10, 320–326. [Google Scholar] [CrossRef]
- Stertz, S.; Reichelt, M.; Spiegel, M.; Kuri, T.; Martinez-Sobrido, L.; Garcia-Sastre, A.; Weber, F.; Kochs, G. The intracellular sites of early replication and budding of SARS-coronavirus. Virology 2007, 361, 304–315. [Google Scholar] [CrossRef] [Green Version]
- Harrison, S.M.; Dove, B.K.; Rothwell, L.; Kaiser, P.; Tarpey, I.; Brooks, G.; Hiscox, J.A. Characterisation of cyclin D1 down-regulation in coronavirus infected cells. Febs. Lett. 2007, 581, 1275–1286. [Google Scholar] [CrossRef] [Green Version]
- Surjit, M.; Liu, B.P.; Jameel, S.; Chow, V.T.K.; Lal, S.K. The SARS coronavirus nucleocapsid protein induces actin reorganization and apoptosis in COS-1 cells in the absence of growth factors. Biochem. J. 2004, 383, 13–18. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhao, G.; Shi, S.Q.; Yang, Y.; Peng, J.P. M and N proteins of SARS coronavirus induce apoptosis in HPF cells. Cell Biol. Toxicol. 2006, 22, 313–322. [Google Scholar] [CrossRef] [PubMed]
- Zhou, B.; Liu, J.L.; Wang, Q.; Liu, X.; Li, X.R.; Li, P.; Ma, Q.J.; Cao, C. The nucleocapsid protein of severe acute respiratory syndrome coronavirus inhibits cell cytokinesis and proliferation by interacting with translation elongation factor 1 alpha. J. Virol. 2008, 82, 6962–6971. [Google Scholar] [CrossRef] [PubMed]
- Hagemeijer, M.C.; Monastyrska, I.; Griffith, J.; van der Sluijs, P.; Voortman, J.; Henegouwen, P.M.V.B.E.; Vonk, A.M.; Rottier, P.J.M.; Reggiori, F.; de Haan, C.A.M. Membrane rearrangements mediated by coronavirus nonstructural proteins 3 and 4. Virology 2014, 458, 125–135. [Google Scholar] [CrossRef] [PubMed]
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Doyle, N.; Hawes, P.C.; Simpson, J.; Adams, L.H.; Maier, H.J. The Porcine Deltacoronavirus Replication Organelle Comprises Double-Membrane Vesicles and Zippered Endoplasmic Reticulum with Double-Membrane Spherules. Viruses 2019, 11, 1030. https://doi.org/10.3390/v11111030
Doyle N, Hawes PC, Simpson J, Adams LH, Maier HJ. The Porcine Deltacoronavirus Replication Organelle Comprises Double-Membrane Vesicles and Zippered Endoplasmic Reticulum with Double-Membrane Spherules. Viruses. 2019; 11(11):1030. https://doi.org/10.3390/v11111030
Chicago/Turabian StyleDoyle, Nicole, Philippa C. Hawes, Jennifer Simpson, Lorin H. Adams, and Helena J. Maier. 2019. "The Porcine Deltacoronavirus Replication Organelle Comprises Double-Membrane Vesicles and Zippered Endoplasmic Reticulum with Double-Membrane Spherules" Viruses 11, no. 11: 1030. https://doi.org/10.3390/v11111030