1887

Abstract

p239 is a virus-like particle constituted from hepatitis E virus (HEV) recombinant proteins. It can be used as a surrogate for HEV and as an investigative tool to study cellular interactions because of its ability to adsorb to and penetrate HepG2 cellular membranes. Our objective was to use p239 to define the role of HEV capsid proteins during the early stages of infection. Pull-down and MALDI-TOF MS experiments identified three host-cell proteins, Grp 78/Bip, -tubulin and heat-shock protein 90 (HSP90), and the latter was investigated further. Antibodies to p239 alone or HSP90 alone could detect p239 or HSP90, suggesting the formation of a complex between p239 and HSP90. In the HepG2 cell, geldanamycin (GA), an HSP90-specific inhibitor, blocked intracellular transportation of p239, but had no effect on the binding and cellular entry of p239, suggesting that HSP90 was important for HEV capsid intracellular transportation. RT-PCR results showed that the efficiency of wild-type HEV infection was inhibited significantly by GA treatment, suggesting the importance of HSP90 in virus infectivity. It was concluded that HSP90 plays a crucial role in the intracellular transportation of viral capsids in the early stage of HEV infection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.019323-0
2010-07-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/91/7/1728.html?itemId=/content/journal/jgv/10.1099/vir.0.019323-0&mimeType=html&fmt=ahah

References

  1. Bradley, D., Andjaparidze, A., Cook, E. H., Jr, McCaustland, K., Balayan, M., Stetler, H., Velazquez, O., Robertson, B., Humphrey, C. & other authors(1988). Aetiological agent of enterically transmitted non-A, non-B hepatitis. J Gen Virol 69, 731–738.[CrossRef] [Google Scholar]
  2. Burch, A. D. & Weller, S. K.(2005). Herpes simplex virus type 1 DNA polymerase requires the mammalian chaperone hsp90 for proper localization to the nucleus. J Virol 79, 10740–10749.[CrossRef] [Google Scholar]
  3. 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] [Google Scholar]
  4. Emerson, S. U., Arankalle, V. A. & Purcell, R. H.(2005). Thermal stability of hepatitis E virus. J Infect Dis 192, 930–933.[CrossRef] [Google Scholar]
  5. Emerson, S. U., Clemente-Casares, P., Moiduddin, N., Arankalle, V. A., Torian, U. & Purcell, R. H.(2006). Putative neutralization epitopes and broad cross-genotype neutralization of hepatitis E virus confirmed by a quantitative cell-culture assay. J Gen Virol 87, 697–704.[CrossRef] [Google Scholar]
  6. 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] [Google Scholar]
  7. Graff, J., Zhou, Y. H., Torian, U., Nguyen, H., St Claire, M., Yu, C., Purcell, R. H. & Emerson, S. U.(2008). Mutations within potential glycosylation sites in the capsid protein of hepatitis E virus prevent the formation of infectious virus particles. J Virol 82, 1185–1194.[CrossRef] [Google Scholar]
  8. Guu, T. S., Liu, Z., Ye, Q., Mata, D. A., Li, K., Yin, C., Zhang, J. & Tao, Y. J.(2009). Structure of the hepatitis E virus-like particle suggests mechanisms for virus assembly and receptor binding. Proc Natl Acad Sci U S A 106, 12992–12997.[CrossRef] [Google Scholar]
  9. Harrell, J. M., Murphy, P. J., Morishima, Y., Chen, H., Mansfield, J. F., Galigniana, M. D. & Pratt, W. B.(2004). Evidence for glucocorticoid receptor transport on microtubules by dynein. J Biol Chem 279, 54647–54654.[CrossRef] [Google Scholar]
  10. He, S., Miao, J., Zheng, Z., Wu, T., Xie, M., Tang, M., Zhang, J., Ng, M. H. & Xia, N.(2008). Putative receptor-binding sites of hepatitis E virus. J Gen Virol 89, 245–249.[CrossRef] [Google Scholar]
  11. Hoffmann, K. & Handschumacher, R. E.(1995). Cyclophilin-40: evidence for a dimeric complex with hsp90. Biochem J 307, 5–8. [Google Scholar]
  12. Hu, J. & Seeger, C.(1996). Hsp90 is required for the activity of a hepatitis B virus reverse transcriptase. Proc Natl Acad Sci U S A 93, 1060–1064.[CrossRef] [Google Scholar]
  13. Hu, J., Flores, D., Toft, D., Wang, X. & Nguyen, D.(2004). Requirement of heat shock protein 90 for human hepatitis B virus reverse transcriptase function. J Virol 78, 13122–13131.[CrossRef] [Google Scholar]
  14. Huang, R., Nakazono, N., Ishii, K., Li, D., Kawamata, O., Kawaguchi, R. & Tsukada, Y.(1995). Hepatitis E virus (87A strain) propagated in A549 cells. J Med Virol 47, 299–302.[CrossRef] [Google Scholar]
  15. Huang, Y. W., Opriessnig, T., Halbur, P. G. & Meng, X. J.(2007). Initiation at the third in-frame AUG codon of open reading frame 3 of the hepatitis E virus is essential for viral infectivity in vivo. J Virol 81, 3018–3026.[CrossRef] [Google Scholar]
  16. Hung, J. J., Chung, C. S. & Chang, W.(2002). Molecular chaperone Hsp90 is important for vaccinia virus growth in cells. J Virol 76, 1379–1390.[CrossRef] [Google Scholar]
  17. Jothikumar, N., Cromeans, T. L., Robertson, B. H., Meng, X. J. & Hill, V. R.(2006). A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. J Virol Methods 131, 65–71.[CrossRef] [Google Scholar]
  18. Kalia, M., Chandra, V., Rahman, S. A., Sehgal, D. & Jameel, S.(2009). Heparan sulfate proteoglycans are required for cellular binding of the hepatitis E virus ORF2 capsid protein and for viral infection. J Virol 83, 12714–12724.[CrossRef] [Google Scholar]
  19. Kim, Y. H., Kim, Y. S., Park, C. H., Chung, I. Y., Yoo, J. M., Kim, J. G., Lee, B. J., Kang, S. S., Cho, G. J. & other authors(2008). Protein kinase C-delta mediates neuronal apoptosis in the retinas of diabetic rats via the Akt signaling pathway. Diabetes 57, 2181–2190.[CrossRef] [Google Scholar]
  20. Kitatsuji, C., Kurogochi, M., Nishimura, S., Ishimori, K. & Wakasugi, K.(2007). Molecular basis of guanine nucleotide dissociation inhibitor activity of human neuroglobin by chemical cross-linking and mass spectrometry. J Mol Biol 368, 150–160.[CrossRef] [Google Scholar]
  21. Kumar, R., Grammatikakis, N. & Chinkers, M.(2001). Regulation of the atrial natriuretic peptide receptor by heat shock protein 90 complexes. J Biol Chem 276, 11371–11375.[CrossRef] [Google Scholar]
  22. Li, S. W., Zhang, J., Li, Y. M., Ou, S. H., Huang, G. Y., He, Z. Q., Ge, S. X., Xian, Y. L., Pang, S. Q. & other authors(2005). A bacterially expressed particulate hepatitis E vaccine: antigenicity, immunogenicity and protectivity on primates. Vaccine 23, 2893–2901.[CrossRef] [Google Scholar]
  23. Li, S., Tang, X., Seetharaman, J., Yang, C., Gu, Y., Zhang, J., Du, H., Shih, J. W., Hew, C. L. & other authors(2009). Dimerization of hepatitis E virus capsid protein E2s domain is essential for virus-host interaction. PLoS Pathog 5, e1000537[CrossRef] [Google Scholar]
  24. Meng, J., Dubreuil, P. & Pillot, J.(1997). A new PCR-based seroneutralization assay in cell culture for diagnosis of hepatitis E. J Clin Microbiol 35, 1373–1377. [Google Scholar]
  25. Meng, J., Dai, X., Chang, J. C., Lopareva, E., Pillot, J., Fields, H. A. & Khudyakov, Y. E.(2001). Identification and characterization of the neutralization epitope(s) of the hepatitis E virus. Virology 288, 203–211.[CrossRef] [Google Scholar]
  26. Mimnaugh, E. G., Xu, W., Vos, M., Yuan, X. & Neckers, L.(2006). Endoplasmic reticulum vacuolization and valosin-containing protein relocalization result from simultaneous hsp90 inhibition by geldanamycin and proteasome inhibition by velcade. Mol Cancer Res 4, 667–681.[CrossRef] [Google Scholar]
  27. Momose, F., Naito, T., Yano, K., Sugimoto, S., Morikawa, Y. & Nagata, K.(2002). Identification of Hsp90 as a stimulatory host factor involved in influenza virus RNA synthesis. J Biol Chem 277, 45306–45314.[CrossRef] [Google Scholar]
  28. Pratt, W. B. & Toft, D. O.(1997). Steroid receptor interactions with heat shock protein and immunophilin chaperones. Endocr Rev 18, 306–360. [Google Scholar]
  29. Pratt, W. B. & Toft, D. O.(2003). Regulation of signaling protein function and trafficking by the hsp90/hsp70-based chaperone machinery. Exp Biol Med (Maywood) 228, 111–133. [Google Scholar]
  30. Reyes-Del Valle, J., Chavez-Salinas, S., Medina, F. & Del Angel, R. M.(2005). Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells. J Virol 79, 4557–4567.[CrossRef] [Google Scholar]
  31. Suuronen, T., Ojala, J., Hyttinen, J. M., Kaarniranta, K., Thornell, A., Kyrylenko, S. & Salminen, A.(2008). Regulation of ER alpha signaling pathway in neuronal HN10 cells: role of protein acetylation and Hsp90. Neurochem Res 33, 1768–1775.[CrossRef] [Google Scholar]
  32. Tanaka, T., Takahashi, M., Kusano, E. & Okamoto, H.(2007). Development and evaluation of an efficient cell-culture system for hepatitis E virus. J Gen Virol 88, 903–911.[CrossRef] [Google Scholar]
  33. Thomas, M., Harrell, J. M., Morishima, Y., Peng, H. M., Pratt, W. B. & Lieberman, A. P.(2006). Pharmacologic and genetic inhibition of hsp90-dependent trafficking reduces aggregation and promotes degradation of the expanded glutamine androgen receptor without stress protein induction. Hum Mol Genet 15, 1876–1883.[CrossRef] [Google Scholar]
  34. Tunon, P. & Johansson, K. E.(1984). Yet another improved silver staining method for the detection of proteins in polyacrylamide gels. J Biochem Biophys Methods 9, 171–179.[CrossRef] [Google Scholar]
  35. Waxman, L., Whitney, M., Pollok, B. A., Kuo, L. C. & Darke, P. L.(2001). Host cell factor requirement for hepatitis C virus enzyme maturation. Proc Natl Acad Sci U S A 98, 13931–13935.[CrossRef] [Google Scholar]
  36. Xia, N. S., Luo, W. X., Zhang, J., Xie, X. Y., Yang, H. J., Li, S. W., Chen, M. & Ng, M. H.(2002). Bioluminescence of Aequorea macrodactyla, a common jellyfish species in the East China Sea. Mar Biotechnol (NY) 4, 155–162.[CrossRef] [Google Scholar]
  37. Yamashita, T., Mori, Y., Miyazaki, N., Cheng, R. H., Yoshimura, M., Unno, H., Shima, R., Moriishi, K., Tsukihara, T. & other authors(2009). Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure. Proc Natl Acad Sci U S A 106, 12986–12991.[CrossRef] [Google Scholar]
  38. Zhang, J., Ge, S. X., Huang, G. Y., Li, S. W., He, Z. Q., Wang, Y. B., Zheng, Y. J., Gu, Y., Ng, M. H. & other authors(2003). Evaluation of antibody-based and nucleic acid-based assays for diagnosis of hepatitis E virus infection in a rhesus monkey model. J Med Virol 71, 518–526.[CrossRef] [Google Scholar]
  39. Zhang, J., Gu, Y., Ge, S. X., Li, S. W., He, Z. Q., Huang, G. Y., Zhuang, H., Ng, M. H. & Xia, N. S.(2005). Analysis of hepatitis E virus neutralization sites using monoclonal antibodies directed against a virus capsid protein. Vaccine 23, 2881–2892.[CrossRef] [Google Scholar]
  40. Zheng, C. F., Simcox, T., Xu, L. & Vaillancourt, P.(1997). A new expression vector for high level protein production, one step purification and direct isotopic labeling of calmodulin-binding peptide fusion proteins. Gene 186, 55–60.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.019323-0
Loading
/content/journal/jgv/10.1099/vir.0.019323-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error