1887

Journal of General Virology header logo

Volume 100, Issue 11

HBx natural variants containing Ser-101 instead of Pro-101 evade ubiquitin-dependent proteasomal degradation by activating proteasomal activator 28 gamma expression Free

Abstract

Proteasomal activator 28 gamma (PA28γ) is frequently overexpressed in hepatocellular carcinoma; however, its underlying mechanism and role in hepatitis B virus (HBV) replication are largely unknown. Here, we found that HBV X protein (HBx) natural variants containing Ser-101 instead of Pro-101 increase reactive oxygen species levels in the mitochondria and activate the ataxia telangiectasia mutated/checkpoint kinase 2 pathway in the nucleus, resulting in the phosphorylation of p53 at Ser-15 and Ser-20 and the subsequent upregulation of its protein levels. Therefore, HBx variants containing Ser-101 induced p53-dependent activation of PA28γ expression in human hepatoma cells. The elevated PA28γ levels upregulated HBx levels through the inhibition of seven in absentia homologue 1-dependent proteasomal degradation. The self-amplifying ability of HBx variants containing Ser-101 via a positive feedback loop involving p53 and PA28γ was accurately reproduced in both a 1.2-mer HBV replicon and HBV infection systems, which also provided evidence for the stimulation of HBV replication by these HBx variants. In conclusion, the ability of HBx to upregulate PA28γ levels via p53 activation, in a Ser-101-dependent pathway, is critical for the stimulation of HBV replication.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): HBx , Hepatitis B virus , p53 , PA28γ and proteasome
© 2019 The Authors
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/content/journal/jgv/10.1099/jgv.0.001337
2019-11-01
2023-10-01
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References

  1. Levrero M, Zucman-Rossi J. Mechanisms of HBV-induced hepatocellular carcinoma. J Hepatol 2016; 64:S84–S101 [View Article]
     [Google Scholar]
  2. Lamontagne RJ, Bagga S, Bouchard MJ. Hepatitis B virus molecular biology and pathogenesis. Hepatoma Res 2016; 2:163–186 [View Article]
     [Google Scholar]
  3. Locarnini S, Zoulim F. Molecular genetics of HBV infection. Antivir Ther 2010; 15:3–14 [View Article]
     [Google Scholar]
  4. Liu S, Koh S, Lee C. Hepatitis B virus X protein and hepatocarcinogenesis. Int J Mol Sci 2016; 17:940 [View Article]
     [Google Scholar]
  5. Tsuge M, Hiraga N, Akiyama R, Tanaka S, Matsushita M et al. Hbx protein is indispensable for development of viraemia in human hepatocyte chimeric mice. J Gen Virol 2010; 91:1854–1864 [View Article]
     [Google Scholar]
  6. Xu Z, Yen TSB, Wu L, Madden CR, Tan W et al. Enhancement of hepatitis B virus replication by its X protein in transgenic mice. J Virol 2002; 76:2579–2584 [View Article]
     [Google Scholar]
  7. Keasler VV, Hodgson AJ, Madden CR, Slagle BL. Hepatitis B virus HBx protein localized to the nucleus restores HBx-deficient virus replication in HepG2 cells and in vivo in hydrodynamically-injected mice. Virology 2009; 390:122–129 [View Article]
     [Google Scholar]
  8. Carmona S, Ely A, Crowther C, Moolla N, Salazar FH et al. Effective inhibition of HBV replication in vivo by anti-HBx short hairpin RNAs. Mol Ther 2006; 13:411–421 [View Article]
     [Google Scholar]
  9. Keasler VV, Hodgson AJ, Madden CR, Slagle BL. Enhancement of hepatitis B virus replication by the regulatory X protein in vitro and in vivo. J Virol 2007; 81:2656–2662 [View Article]
     [Google Scholar]
  10. Henkler F, Hoare J, Waseem N, Goldin RD, McGarvey MJ et al. Intracellular localization of the hepatitis B virus HBx protein. J Gen Virol 2001; 82:871–882 [View Article]
     [Google Scholar]
  11. Cha MY, Ryu DK, Jung HS, Chang HE, Ryu WS. Stimulation of hepatitis B virus genome replication by HBx is linked to both nuclear and cytoplasmic HBx expression. J Gen Virol 2009; 90:978–986 [View Article]
     [Google Scholar]
  12. Huh KW, Siddiqui A. Characterization of the mitochondrial association of hepatitis B virus X protein, HBx. Mitochondrion 2002; 1:349–359 [View Article]
     [Google Scholar]
  13. Tang H, Delgermaa L, Huang F, Oishi N, Liu L et al. The transcriptional transactivation function of HBx protein is important for its augmentation role in hepatitis B virus replication. J Virol 2005; 79:5548–5556 [View Article]
     [Google Scholar]
  14. Bouchard MJ, Wang LH, Schneider RJ. Calcium signaling by HBx protein in hepatitis B virus DNA replication. Science 2001; 294:2376–2378 [View Article]
     [Google Scholar]
  15. Rawat S, Bouchard MJ. The hepatitis B virus (HBV) HBx protein activates Akt to simultaneously regulate HBV replication and hepatocyte survival. J Virol 2015; 89:999–1012 [View Article]
     [Google Scholar]
  16. Coux O, Tanaka K, Goldberg AL. Structure and functions of the 20S and 26S proteasomes. Annu Rev Biochem 1996; 65:801–847 [View Article]
     [Google Scholar]
  17. Mao I, Liu J, Li X, Luo H, REGgamma LH. Reggamma, a proteasome activator and beyond?. Cell Mol Life Sci 2008; 65:3971–3980 [View Article]
     [Google Scholar]
  18. Chen X, Barton LF, Chi Y, Clurman BE, Roberts JM. Ubiquitin-Independent degradation of cell-cycle inhibitors by the REGgamma proteasome. Mol Cell 2007; 26:843–852 [View Article]
     [Google Scholar]
  19. Li X, Amazit L, Long W, Lonard DM, Monaco JJ et al. Ubiquitin- and ATP-independent proteolytic turnover of p21 by the REGgamma-proteasome pathway. Mol Cell 2007; 26:831–842 [View Article]
     [Google Scholar]
  20. Liu J, Yu G, Zhao Y, Zhao D, Wang Y et al. Reggamma modulates p53 activity by regulating its cellular localization. J Cell Sci 2010; 123:4076–4084 [View Article]
     [Google Scholar]
  21. Zhang Z, Zhang R. Proteasome activator PA28 gamma regulates p53 by enhancing its Mdm2-mediated degradation. Embo J 2008; 27:852–864 [View Article]
     [Google Scholar]
  22. Yu G, Zhao Y, He J, Lonard DM, Mao CA et al. Comparative analysis of REG{gamma} expression in mouse and human tissues. J Mol Cell Biol 2010; 2:192–198 [View Article]
     [Google Scholar]
  23. Wan ZX, Yuan DM, Zhuo YM, Yi X, Zhou J et al. The proteasome activator PA28γ, a negative regulator of p53, is transcriptionally up-regulated by p53. Int J Mol Sci 2014; 15:2573–2584 [View Article]
     [Google Scholar]
  24. He J, Cui L, Zeng Y, Wang G, Zhou P et al. Regγ is associated with multiple oncogenic pathways in human cancers. BMC Cancer 2012; 12:75 [View Article]
     [Google Scholar]
  25. Yeom S, Jeong H, Kim SS, Jang KL. Hepatitis B virus X protein activates proteasomal activator 28 gamma expression via upregulation of p53 levels to stimulate virus replication. J Gen Virol 2018; 99:655–666 [View Article]
     [Google Scholar]
  26. Kim S, Lee HS, Ji JH, Cho MY, Yoo YS et al. Hepatitis B virus X protein activates the ATM-Chk2 pathway and delays cell cycle progression. J Gen Virol 2015; 96:2242–2251 [View Article]
     [Google Scholar]
  27. Yeom S, Kim SS, Jeong H, Jang KL. Hepatitis B virus X protein activates E3 ubiquitin ligase Siah-1 to control virus propagation via a negative feedback loop. J Gen Virol 2017; 98:1774–1784 [View Article]
     [Google Scholar]
  28. Wang WH, Grégori G, Hullinger RL, Andrisani OM. Sustained activation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase pathways by hepatitis B virus X protein mediates apoptosis via induction of Fas/FasL and tumor necrosis factor (TNF) receptor 1/TNF-alpha expression. Mol Cell Biol 2004; 24:10352–10365 [View Article]
     [Google Scholar]
  29. Kwun HJ, Jang KL. Natural variants of hepatitis B virus X protein have differential effects on the expression of cyclin-dependent kinase inhibitor p21 gene. Nucleic Acids Res 2004; 32:2202–2213 [View Article]
     [Google Scholar]
  30. Lee YI, Hwang JM, Im JH, Lee YI, Kim NS et al. Human hepatitis B virus-X protein alters mitochondrial function and physiology in human liver cells. J Biol Chem 2004; 279:15460–15471 [View Article]
     [Google Scholar]
  31. Matsuda Y, Sanpei A, Wakai T, Kubota M, Osawa M et al. Hepatitis B virus X stimulates redox signaling through activation of ataxia telangiectasia mutated kinase. Int J Clin Exp Pathol 2014; 7:2032–2043
     [Google Scholar]
  32. Fiucci G, Beaucourt S, Duflaut D, Lespagnol A, Stumptner-Cuvelette P et al. Siah-1b is a direct transcriptional target of p53: identification of the functional p53 responsive element in the siah-1b promoter. Proc Natl Acad Sci USA 2004; 101:3510–3515 [View Article]
     [Google Scholar]
  33. Zhao J, Wang C, Wang J, Yang X, Diao N et al. E3 ubiquitin ligase Siah-1 facilitates poly-ubiquitylation and proteasomal degradation of the hepatitis B viral X protein. FEBS Lett 2011; 585:2943–2950 [View Article]
     [Google Scholar]
  34. Cha MY, Kim CM, Park YM, Ryu WS. Hepatitis B virus X protein is essential for the activation of Wnt/beta-catenin signaling in hepatoma cells. Hepatology 2004; 39:1683–1693 [View Article]
     [Google Scholar]
  35. Elmore LW, Hancock AR, Chang SF, Wang XW, Chang S et al. Hepatitis B virus X protein and p53 tumor suppressor interactions in the modulation of apoptosis. Proc Natl Acad Sci USA 1997; 94:14707–14712 [View Article]
     [Google Scholar]
  36. Wang XW, Gibson MK, Vermeulen W, Yeh H, Forrester K et al. Abrogation of p53-induced apoptosis by the hepatitis B virus X gene. Cancer Res 1995; 55:6012–6016
     [Google Scholar]
  37. Feitelson MA, Zhu M, Duan LX, London WT. Hepatitis B X antigen and p53 are associated in vitro and in liver tissues from patients with primary hepatocellular carcinoma. Oncogene 1993; 8:1109–1117
     [Google Scholar]
  38. Wang XW, Forrester K, Yeh H, Feitelson MA, Gu JR et al. Hepatitis B virus X protein inhibits p53 sequence-specific DNA binding, transcriptional activity, and association with transcription factor ERCC3. Proc Natl Acad Sci USA 1994; 91:2230–2234 [View Article]
     [Google Scholar]
  39. Ahn JY, Jung EY, Kwun HJ, Lee CW, Sung YC et al. Dual effects of hepatitis B virus X protein on the regulation of cell-cycle control depending on the status of cellular p53. J Gen Virol 2002; 83:2765–2772 [View Article]
     [Google Scholar]
  40. Jung JK, Kwun HJ, Lee JO, Arora P, Jang KL. Hepatitis B virus X protein differentially affects the ubiquitin-mediated proteasomal degradation of beta-catenin depending on the status of cellular p53. J Gen Virol 2007; 88:2144–2154 [View Article]
     [Google Scholar]
  41. Wang WH, Hullinger RL, Andrisani OM. Hepatitis B virus X protein via the p38MAPK pathway induces E2F1 release and ATR kinase activation mediating p53 apoptosis. J Biol Chem 2008; 283:25455–25467 [View Article]
     [Google Scholar]
  42. Knoll S, Fürst K, Thomas S, Villanueva Baselga S, Stoll A et al. Dissection of cell context-dependent interactions between HBx and p53 family members in regulation of apoptosis: a role for HBV-induced HCC. Cell Cycle 2011; 10:3554–3565 [View Article]
     [Google Scholar]
  43. Blum HE, Zhang ZS, Galun E, von Weizsäcker F, Garner B et al. Hepatitis B virus X protein is not central to the viral life cycle in vitro. J Virol 1992; 66:1223–1227
     [Google Scholar]
  44. Lee CW, Sørensen TS, Shikama N, La Thangue NB. Functional interplay between p53 and E2F through co-activator p300. Oncogene 1998; 16:2695–2710 [View Article]
     [Google Scholar]
  45. Kwak J, Tiwari I, Jang KL. Hepatitis C virus core activates proteasomal activator 28γ expression via upregulation of p53 levels to control virus propagation. J Gen Virol 2017; 98:56–67 [View Article]
     [Google Scholar]
  46. Mendy ME, Kaye S, van der Sande M, Rayco-Solon P, Waight PA et al. Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in the Gambia. Virol J 2006; 3:23 [View Article]
     [Google Scholar]
  47. Severi T, Vander Borght S, Libbrecht L, VanAelst L, Nevens F et al. Hbx or HCV core gene expression in HepG2 human liver cells results in a survival benefit against oxidative stress with possible implications for HCC development. Chem Biol Interact 2007; 168:128–134 [View Article]
     [Google Scholar]
  48. Lim JS, Park SH, Jang KL. Hepatitis C virus core protein overcomes stress-induced premature senescence by down-regulating p16 expression via DNA methylation. Cancer Lett 2012; 321:154–161 [View Article]
     [Google Scholar]
  49. Lee JO, Kwun HJ, Jung JK, Choi KH, Min DS et al. Hepatitis B virus X protein represses E-cadherin expression via activation of DNA methyltransferase 1. Oncogene 2005; 24:6617–6625 [View Article]
     [Google Scholar]
  50. Ponchel F, Toomes C, Bransfield K, Leong FT, Douglas SH et al. Real-Time PCR based on SYBR-Green I fluorescence: an alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. BMC Biotechnol 2003; 3:18 [View Article]
     [Google Scholar]
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HBx natural variants containing Ser-101 instead of Pro-101 evade ubiquitin-dependent proteasomal degradation by activating proteasomal activator 28 gamma expression
J. Gen. Virol. 100, 1554 (2019); https://doi.org/10.1099/jgv.0.001337
/content/journal/jgv/10.1099/jgv.0.001337
/content/journal/jgv/10.1099/jgv.0.001337
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