1887

Abstract

Hepatitis E virus (HEV) is a positive-strand RNA virus and a major causative agent of acute sporadic and epidemic hepatitis. HEV replication protein is encoded by ORF1 and contains the predicted domains of methyltransferase (MT), protease, macro domain, helicase (HEL) and polymerase (POL). In this study, the full-length protein pORF1 (1693 aa) and six truncated variants were expressed by translation and in human HeLa and hepatic Huh-7 cells by using several vector systems. The proteins were visualized by three specific antisera directed against the MT, HEL and POL domains. translation of full-length pORF1 yielded smaller quantities of two fragments. However, these fragments were not observed after pORF1 expression and pulse–chase studies in human cells, and their production was not dependent on the predicted protease domain in pORF1. The weight of evidence supports the proposition that pORF1 is not subjected to specific proteolytic processing, which is unusual among animal positive-strand RNA viruses but common for plant viruses. pORF1 was membrane associated in cells and localized to a perinuclear region, where it partially overlapped with localization of the endoplasmic reticulum (ER) marker BAP31 and was closely interspersed with staining of the ER–Golgi intermediate compartment marker protein ERGIC-53. Co-localization with BAP31 was enhanced by treatment with brefeldin A. Therefore, HEV may utilize modified early secretory pathway membranes for replication.

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2013-04-01
2019-10-21
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References

  1. Agrawal S., Gupta D., Panda S. K.. ( 2001;). The 3′ end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp). . Virology 282:, 87–101. [CrossRef][PubMed]
    [Google Scholar]
  2. Ahmad I., Holla R. P., Jameel S.. ( 2011;). Molecular virology of hepatitis E virus. . Virus Res 161:, 47–58. [CrossRef][PubMed]
    [Google Scholar]
  3. Ahola T., Kääriäinen L.. ( 1995;). Reaction in alphavirus mRNA capping: formation of a covalent complex of nonstructural protein nsP1 with 7-methyl-GMP. . Proc Natl Acad Sci U S A 92:, 507–511. [CrossRef][PubMed]
    [Google Scholar]
  4. Ahola T., Lampio A., Auvinen P., Kääriäinen L.. ( 1999;). Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity. . EMBO J 18:, 3164–3172. [CrossRef][PubMed]
    [Google Scholar]
  5. Ansari I. H., Nanda S. K., Durgapal H., Agrawal S., Mohanty S. K., Gupta D., Jameel S., Panda S. K.. ( 2000;). Cloning, sequencing, and expression of the hepatitis E virus (HEV) nonstructural open reading frame 1 (ORF1). . J Med Virol 60:, 275–283. [CrossRef][PubMed]
    [Google Scholar]
  6. Bolte S., Cordelières F. P.. ( 2006;). A guided tour into subcellular colocalization analysis in light microscopy. . J Microsc 224:, 213–232. [CrossRef][PubMed]
    [Google Scholar]
  7. Chikkanna-Gowda C. P., Sheahan B. J., Fleeton M. N., Atkins G. J.. ( 2005;). Regression of mouse tumours and inhibition of metastases following administration of a Semliki Forest virus vector with enhanced expression of IL-12. . Gene Ther 12:, 1253–1263. [CrossRef][PubMed]
    [Google Scholar]
  8. Emerson S. U., Nguyen H., Graff J., Stephany D. A., Brockington A., Purcell R. H.. ( 2004;). In vitro replication of hepatitis E virus (HEV) genomes and of an HEV replicon expressing green fluorescent protein. . J Virol 78:, 4838–4846. [CrossRef][PubMed]
    [Google Scholar]
  9. Emerson S. U., Nguyen H. T., Torian U., Burke D., Engle R., Purcell R. H.. ( 2010;). Release of genotype 1 hepatitis E virus from cultured hepatoma and polarized intestinal cells depends on open reading frame 3 protein and requires an intact PXXP motif. . J Virol 84:, 9059–9069. [CrossRef][PubMed]
    [Google Scholar]
  10. Graff J., Torian U., Nguyen H., Emerson S. U.. ( 2006;). A bicistronic subgenomic mRNA encodes both the ORF2 and ORF3 proteins of hepatitis E virus. . J Virol 80:, 5919–5926. [CrossRef][PubMed]
    [Google Scholar]
  11. Hauri H. P., Kappeler F., Andersson H., Appenzeller C.. ( 2000;). ERGIC-53 and traffic in the secretory pathway. . J Cell Sci 113:, 587–596.[PubMed]
    [Google Scholar]
  12. Hsu N.-Y., Ilnytska O., Belov G., Santiana M., Chen Y.-H., Takvorian P. M., Pau C., van der Schaar H., Kaushik-Basu N.. & other authors ( 2010;). Viral reorganization of the secretory pathway generates distinct organelles for RNA replication. . Cell 141:, 799–811. [CrossRef][PubMed]
    [Google Scholar]
  13. Karpe Y. A., Lole K. S.. ( 2010a;). NTPase and 5′ to 3′ RNA duplex-unwinding activities of the hepatitis E virus helicase domain. . J Virol 84:, 3595–3602. [CrossRef][PubMed]
    [Google Scholar]
  14. Karpe Y. A., Lole K. S.. ( 2010b;). RNA 5′-triphosphatase activity of the hepatitis E virus helicase domain. . J Virol 84:, 9637–9641. [CrossRef][PubMed]
    [Google Scholar]
  15. Karpe Y. A., Lole K. S.. ( 2011;). Deubiquitination activity associated with hepatitis E virus putative papain-like cysteine protease. . J Gen Virol 92:, 2088–2092. [CrossRef][PubMed]
    [Google Scholar]
  16. Klausner R. D., Donaldson J. G., Lippincott-Schwartz J.. ( 1992;). Brefeldin A: insights into the control of membrane traffic and organelle structure. . J Cell Biol 116:, 1071–1080. [CrossRef][PubMed]
    [Google Scholar]
  17. Koonin E. V., Gorbalenya A. E., Purdy M. A., Rozanov M. N., Reyes G. R., Bradley D. W.. ( 1992;). Computer-assisted assignment of functional domains in the nonstructural polyprotein of hepatitis E virus: delineation of an additional group of positive-strand RNA plant and animal viruses. . Proc Natl Acad Sci U S A 89:, 8259–8263. [CrossRef][PubMed]
    [Google Scholar]
  18. Kujala P., Ahola T., Ehsani N., Auvinen P., Vihinen H., Kääriäinen L.. ( 1999;). Intracellular distribution of rubella virus nonstructural protein P150. . J Virol 73:, 7805–7811.[PubMed]
    [Google Scholar]
  19. Liu X., Ropp S. L., Jackson R. J., Frey T. K.. ( 1998;). The rubella virus nonstructural protease requires divalent cations for activity and functions in trans. . J Virol 72:, 4463–4466.[PubMed]
    [Google Scholar]
  20. Määttä J., Hallikas O., Welti S., Hildén P., Schröder J., Kuismanen E.. ( 2000;). Limited caspase cleavage of human BAP31. . FEBS Lett 484:, 202–206. [CrossRef][PubMed]
    [Google Scholar]
  21. Magden J., Takeda N., Li T., Auvinen P., Ahola T., Miyamura T., Merits A., Kääriäinen L.. ( 2001;). Virus-specific mRNA capping enzyme encoded by hepatitis E virus. . J Virol 75:, 6249–6255. [CrossRef][PubMed]
    [Google Scholar]
  22. Mori Y., Matsuura Y.. ( 2011;). Structure of hepatitis E viral particle. . Virus Res 161:, 59–64. [CrossRef][PubMed]
    [Google Scholar]
  23. Neuvonen M., Ahola T.. ( 2009;). Differential activities of cellular and viral macro domain proteins in binding of ADP-ribose metabolites. . J Mol Biol 385:, 212–225. [CrossRef][PubMed]
    [Google Scholar]
  24. Okamoto H.. ( 2011;). Hepatitis E virus cell culture models. . Virus Res 161:, 65–77. [CrossRef][PubMed]
    [Google Scholar]
  25. Panda S. K., Ansari I. H., Durgapal H., Agrawal S., Jameel S.. ( 2000;). The in vitro-synthesized RNA from a cDNA clone of hepatitis E virus is infectious. . J Virol 74:, 2430–2437. [CrossRef][PubMed]
    [Google Scholar]
  26. Purcell R. H., Emerson S. U.. ( 2008;). Hepatitis E: an emerging awareness of an old disease. . J Hepatol 48:, 494–503. [CrossRef][PubMed]
    [Google Scholar]
  27. Reggiori F., Monastyrska I., Verheije M. H., Calì T., Ulasli M., Bianchi S., Bernasconi R., de Haan C. A., Molinari M.. ( 2010;). Coronaviruses hijack the LC3-I-positive EDEMosomes, ER-derived vesicles exporting short-lived ERAD regulators, for replication. . Cell Host Microbe 7:, 500–508. [CrossRef][PubMed]
    [Google Scholar]
  28. Ropp S. L., Tam A. W., Beames B., Purdy M., Frey T. K.. ( 2000;). Expression of the hepatitis E virus ORF1. . Arch Virol 145:, 1321–1337. [CrossRef][PubMed]
    [Google Scholar]
  29. Salonen A., Vasiljeva L., Merits A., Magden J., Jokitalo E., Kääriäinen L.. ( 2003;). Properly folded nonstructural polyprotein directs the Semliki Forest virus replication complex to the endosomal compartment. . J Virol 77:, 1691–1702. [CrossRef][PubMed]
    [Google Scholar]
  30. Salonen A., Ahola T., Kääriäinen L.. ( 2005;). Viral RNA replication in association with cellular membranes. . Curr Top Microbiol Immunol 285:, 139–173. [CrossRef][PubMed]
    [Google Scholar]
  31. Sancho M. C., Schleich S., Griffiths G., Krijnse-Locker J.. ( 2002;). The block in assembly of modified vaccinia virus Ankara in HeLa cells reveals new insights into vaccinia virus morphogenesis. . J Virol 76:, 8318–8334. [CrossRef][PubMed]
    [Google Scholar]
  32. Saraste J., Svensson K.. ( 1991;). Distribution of the intermediate elements operating in ER to Golgi transport. . J Cell Sci 100:, 415–430.[PubMed]
    [Google Scholar]
  33. Sehgal D., Thomas S., Chakraborty M., Jameel S.. ( 2006;). Expression and processing of the hepatitis E virus ORF1 nonstructural polyprotein. . Virol J 3:, 38. [CrossRef][PubMed]
    [Google Scholar]
  34. Spuul P., Balistreri G., Hellström K., Golubtsov A. V., Jokitalo E., Ahola T.. ( 2011;). Assembly of alphavirus replication complexes from RNA and protein components in a novel trans-replication system in mammalian cells. . J Virol 85:, 4739–4751. [CrossRef][PubMed]
    [Google Scholar]
  35. Suppiah S., Zhou Y., Frey T. K.. ( 2011;). Lack of processing of the expressed ORF1 gene product of hepatitis E virus. . Virol J 8:, 245. [CrossRef][PubMed]
    [Google Scholar]
  36. Thakral D., Nayak B., Rehman S., Durgapal H., Panda S. K.. ( 2005;). Replication of a recombinant hepatitis E virus genome tagged with reporter genes and generation of a short-term cell line producing viral RNA and proteins. . J Gen Virol 86:, 1189–1200. [CrossRef][PubMed]
    [Google Scholar]
  37. van der Heijden M. W., Bol J. F.. ( 2002;). Composition of alphavirus-like replication complexes: involvement of virus and host encoded proteins. . Arch Virol 147:, 875–898. [CrossRef][PubMed]
    [Google Scholar]
  38. Vasiljeva L., Merits A., Golubtsov A., Sizemskaja V., Kääriäinen L., Ahola T.. ( 2003;). Regulation of the sequential processing of Semliki Forest virus replicase polyprotein. . J Biol Chem 278:, 41636–41645. [CrossRef][PubMed]
    [Google Scholar]
  39. Wyatt L. S., Moss B., Rozenblatt S.. ( 1995;). Replication-deficient vaccinia virus encoding bacteriophage T7 RNA polymerase for transient gene expression in mammalian cells. . Virology 210:, 202–205. [CrossRef][PubMed]
    [Google Scholar]
  40. Yamada K., Takahashi M., Hoshino Y., Takahashi H., Ichiyama K., Nagashima S., Tanaka T., Okamoto H.. ( 2009a;). ORF3 protein of hepatitis E virus is essential for virion release from infected cells. . J Gen Virol 90:, 1880–1891. [CrossRef][PubMed]
    [Google Scholar]
  41. Yamada K., Takahashi M., Hoshino Y., Takahashi H., Ichiyama K., Tanaka T., Okamoto H.. ( 2009b;). Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells. . J Gen Virol 90:, 457–462. [CrossRef][PubMed]
    [Google Scholar]
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