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Journal of General Virology header logo Journal of General Virology

Volume 103, Issue 4

Activities of endogenous APOBEC3s and uracil-DNA-glycosylase affect the hypermutation frequency of hepatitis B virus cccDNA Open Access

Abstract

The covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) plays a key role in the persistence of viral infection. We have previously shown that overexpression of an antiviral factor APOBEC3G (A3G) induces hypermutation in duck HBV (DHBV) cccDNA, whereas uracil-DNA-glycosylase (UNG) reduces these mutations. In this study, using cell-culture systems, we examined whether endogenous A3s and UNG affect HBV cccDNA mutation frequency. IFNγ stimulation induced a significant increase in endogenous A3G expression and cccDNA hypermutation. UNG inhibition enhanced the IFNγ-mediated hypermutation frequency. Transfection of reconstructed cccDNA revealed that this enhanced hypermutation caused a reduction in viral replication. These results suggest that the balance of endogenous A3s and UNG activities affects HBV cccDNA mutation and replication competency.

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  • Accepted:
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Keyword(s): APOBEC , DNA repair , hepatitis B virus , host factor and mutation
Funding
This study was supported by the:
  • Takeda Science Foundation
    • Principle Award Recipient: KouichiKitamura
  • Japan Agency for Medical Research and Development (Award JP19fk0210053)
    • Principle Award Recipient: MasamichiMuramatsu
  • Japan Agency for Medical Research and Development (Award JP18fk0310103)
    • Principle Award Recipient: MasamichiMuramatsu
  • Japan Agency for Medical Research and Development (Award JP18fk0310119)
    • Principle Award Recipient: KouichiKitamura
  • Japan Society for the Promotion of Science (Award 19K07583)
    • Principle Award Recipient: MasamichiMuramatsu
  • Japan Society for the Promotion of Science (Award 20K08380)
    • Principle Award Recipient: KouichiKitamura
© 2022 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2022-04-19
2025-05-16
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References

  1. Trépo C, Chan HLY, Lok A. Hepatitis B virus infection. Lancet 2014; 384:2053–2063 [View Article] [PubMed]
     [Google Scholar]
  2. Seeger C, Mason WS. Molecular biology of hepatitis B virus infection. Virology 2015; 479–480:672–686 [View Article] [PubMed]
     [Google Scholar]
  3. Harris RS, Dudley JP. APOBECs and virus restriction. Virology 2015; 479–480:131–145 [View Article] [PubMed]
     [Google Scholar]
  4. Rösler C, Köck J, Malim MH, Blum HE, von Weizsäcker F. Comment on “Inhibition of hepatitis B virus replication by APOBEC3G.”. Science 2004; 305:1403; author reply 1403 [View Article] [PubMed]
     [Google Scholar]
  5. Suspène R, Guétard D, Henry M, Sommer P, Wain-Hobson S et al. Extensive editing of both hepatitis B virus DNA strands by APOBEC3 cytidine deaminases in vitro and in vivo. Proc Natl Acad Sci U S A 2005; 102:8321–8326 [View Article] [PubMed]
     [Google Scholar]
  6. Jost S, Turelli P, Mangeat B, Protzer U, Trono D. Induction of antiviral cytidine deaminases does not explain the inhibition of hepatitis B virus replication by interferons. J Virol 2007; 81:10588–10596 [View Article] [PubMed]
     [Google Scholar]
  7. Nguyen DH, Gummuluru S, Hu J. Deamination-independent inhibition of hepatitis B virus reverse transcription by APOBEC3G. J Virol 2007; 81:4465–4472 [View Article] [PubMed]
     [Google Scholar]
  8. Kitamura K, Wang Z, Chowdhury S, Simadu M, Koura M et al. Uracil DNA glycosylase counteracts APOBEC3G-induced hypermutation of hepatitis B viral genomes: excision repair of covalently closed circular DNA. PLoS Pathog 2013; 9:e1003361 [View Article] [PubMed]
     [Google Scholar]
  9. Watashi K, Liang G, Iwamoto M, Marusawa H, Uchida N et al. Interleukin-1 and tumor necrosis factor-α trigger restriction of hepatitis B virus infection via a cytidine deaminase activation-induced cytidine deaminase (AID). J Biol Chem 2013; 288:31715–31727 [View Article] [PubMed]
     [Google Scholar]
  10. Lucifora J, Xia Y, Reisinger F, Zhang K, Stadler D et al. Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA. Science 2014; 343:1221–1228 [View Article] [PubMed]
     [Google Scholar]
  11. Xia Y, Stadler D, Lucifora J, Reisinger F, Webb D et al. Interferon-γ and Tumor Necrosis Factor-α Produced by T Cells Reduce the HBV Persistence Form, cccDNA, Without Cytolysis. Gastroenterology 2016; 150:194–205 [View Article] [PubMed]
     [Google Scholar]
  12. Bockmann J-H, Stadler D, Xia Y, Ko C, Wettengel JM et al. Comparative Analysis of the Antiviral Effects Mediated by Type I and III Interferons in Hepatitis B Virus-Infected Hepatocytes. J Infect Dis 2019; 220:567–577 [View Article] [PubMed]
     [Google Scholar]
  13. Guidotti LG, Ando K, Hobbs MV, Ishikawa T, Runkel L et al. Cytotoxic T lymphocytes inhibit hepatitis B virus gene expression by a noncytolytic mechanism in transgenic mice. Proc Natl Acad Sci U S A 1994; 91:3764–3768 [View Article] [PubMed]
     [Google Scholar]
  14. Guidotti LG, Ishikawa T, Hobbs MV, Matzke B, Schreiber R et al. Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity 1996; 4:25–36 [View Article] [PubMed]
     [Google Scholar]
  15. Guidotti LG, Rochford R, Chung J, Shapiro M, Purcell R et al. Viral clearance without destruction of infected cells during acute HBV infection. Science 1999; 284:825–829 [View Article] [PubMed]
     [Google Scholar]
  16. Thimme R, Wieland S, Steiger C, Ghrayeb J, Reimann KA et al. CD8(+) T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection. J Virol 2003; 77:68–76 [View Article] [PubMed]
     [Google Scholar]
  17. Phillips S, Chokshi S, Riva A, Evans A, Williams R et al. CD8(+) T cell control of hepatitis B virus replication: direct comparison between cytolytic and noncytolytic functions. J Immunol 2010; 184:287–295 [View Article] [PubMed]
     [Google Scholar]
  18. Vartanian J-P, Henry M, Marchio A, Suspène R, Aynaud M-M et al. Massive APOBEC3 editing of hepatitis B viral DNA in cirrhosis. PLoS Pathog 2010; 6:e1000928 [View Article] [PubMed]
     [Google Scholar]
  19. Sousa MML, Krokan HE, Slupphaug G. DNA-uracil and human pathology. Mol Aspects Med 2007; 28:276–306 [View Article] [PubMed]
     [Google Scholar]
  20. Chowdhury S, Kitamura K, Simadu M, Koura M, Muramatsu M. Concerted action of activation-induced cytidine deaminase and uracil-DNA glycosylase reduces covalently closed circular DNA of duck hepatitis B virus. FEBS Lett 2013; 587:3148–3152 [View Article] [PubMed]
     [Google Scholar]
  21. Stadler D, Kächele M, Jones AN, Hess J, Urban C et al. Interferon-induced degradation of the persistent hepatitis B virus cccDNA form depends on ISG20. EMBO Rep 2021; 22:e49568 [View Article] [PubMed]
     [Google Scholar]
  22. Hagen L, Kavli B, Sousa MML, Torseth K, Liabakk NB et al. Cell cycle-specific UNG2 phosphorylations regulate protein turnover, activity and association with RPA. EMBO J 2008; 27:51–61 [View Article] [PubMed]
     [Google Scholar]
  23. Sanderson RJ, Mosbaugh DW. Identification of specific carboxyl groups on uracil-DNA glycosylase inhibitor protein that are required for activity. J Biol Chem 1996; 271:29170–29181 [View Article] [PubMed]
     [Google Scholar]
  24. Begum NA, Kinoshita K, Kakazu N, Muramatsu M, Nagaoka H et al. Uracil DNA glycosylase activity is dispensable for immunoglobulin class switch. Science 2004; 305:1160–1163 [View Article] [PubMed]
     [Google Scholar]
  25. Wang Z, Wakae K, Kitamura K, Aoyama S, Liu G et al. APOBEC3 deaminases induce hypermutation in human papillomavirus 16 DNA upon beta interferon stimulation. J Virol 2014; 88:1308–1317 [View Article] [PubMed]
     [Google Scholar]
  26. Zhou W, Ma Y, Zhang J, Hu J, Zhang M et al. Predictive model for inflammation grades of chronic hepatitis B: Large-scale analysis of clinical parameters and gene expressions. Liver Int 2017; 37:1632–1641 [View Article] [PubMed]
     [Google Scholar]
  27. Li Y, Que L, Fukano K, Koura M, Kitamura K et al. MCPIP1 reduces HBV-RNA by targeting its epsilon structure. Sci Rep 2020; 10:20763 [View Article] [PubMed]
     [Google Scholar]
  28. Iwamoto M, Cai D, Sugiyama M, Suzuki R, Aizaki H et al. Functional association of cellular microtubules with viral capsid assembly supports efficient hepatitis B virus replication. Sci Rep 2017; 7:10620 [View Article] [PubMed]
     [Google Scholar]
  29. Sells MA, Chen ML, Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A 1987; 84:1005–1009 [View Article] [PubMed]
     [Google Scholar]
  30. Iwamoto M, Watashi K, Tsukuda S, Aly HH, Fukasawa M et al. Evaluation and identification of hepatitis B virus entry inhibitors using HepG2 cells overexpressing a membrane transporter NTCP. Biochem Biophys Res Commun 2014; 443:808–813 [View Article] [PubMed]
     [Google Scholar]
  31. Liang G, Liu G, Kitamura K, Wang Z, Chowdhury S et al. TGF-β suppression of HBV RNA through AID-dependent recruitment of an RNA exosome complex. PLoS Pathog 2015; 11:e1004780 [View Article] [PubMed]
     [Google Scholar]
  32. Kitamura K, Que L, Shimadu M, Koura M, Ishihara Y et al. Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus. PLoS Pathog 2018; 14:e1007124 [View Article] [PubMed]
     [Google Scholar]
  33. Margeridon S, Carrouée-Durantel S, Chemin I, Barraud L, Zoulim F et al. Rolling circle amplification, a powerful tool for genetic and functional studies of complete hepatitis B virus genomes from low-level infections and for directly probing covalently closed circular DNA. Antimicrob Agents Chemother 2008; 52:3068–3073 [View Article] [PubMed]
     [Google Scholar]
  34. Bonvin M, Achermann F, Greeve I, Stroka D, Keogh A et al. Interferon-inducible expression of APOBEC3 editing enzymes in human hepatocytes and inhibition of hepatitis B virus replication. Hepatology 2006; 43:1364–1374 [View Article] [PubMed]
     [Google Scholar]
  35. Kanagaraj A, Sakamoto N, Que L, Li Y, Mohiuddin M et al. Different antiviral activities of natural APOBEC3C, APOBEC3G, and APOBEC3H variants against hepatitis B virus. Biochem Biophys Res Commun 2019; 518:26–31 [View Article] [PubMed]
     [Google Scholar]
  36. Chelico L, Pham P, Calabrese P, Goodman MF. APOBEC3G DNA deaminase acts processively 3’ --> 5’ on single-stranded DNA. Nat Struct Mol Biol 2006; 13:392–399 [View Article] [PubMed]
     [Google Scholar]
  37. Königer C, Wingert I, Marsmann M, Rösler C, Beck J et al. Involvement of the host DNA-repair enzyme TDP2 in formation of the covalently closed circular DNA persistence reservoir of hepatitis B viruses. Proc Natl Acad Sci U S A 2014; 111:E4244–53 [View Article] [PubMed]
     [Google Scholar]
  38. Cui X, McAllister R, Boregowda R, Sohn JA, Cortes Ledesma F et al. Does Tyrosyl DNA Phosphodiesterase-2 Play a Role in Hepatitis B Virus Genome Repair?. PLoS One 2015; 10:e0128401 [View Article] [PubMed]
     [Google Scholar]
  39. Qi Y, Gao Z, Xu G, Peng B, Liu C et al. DNA Polymerase κ Is a Key Cellular Factor for the Formation of Covalently Closed Circular DNA of Hepatitis B Virus. PLoS Pathog 2016; 12:e1005893 [View Article] [PubMed]
     [Google Scholar]
  40. Long Q, Yan R, Hu J, Cai D, Mitra B et al. The role of host DNA ligases in hepadnavirus covalently closed circular DNA formation. PLoS Pathog 2017; 13:e1006784 [View Article] [PubMed]
     [Google Scholar]
  41. Sheraz M, Cheng J, Tang L, Chang J, Guo J-T. Cellular DNA Topoisomerases Are Required for the Synthesis of Hepatitis B Virus Covalently Closed Circular DNA. J Virol 2019; 93:e02230-18 [View Article] [PubMed]
     [Google Scholar]
  42. Tang L, Sheraz M, McGrane M, Chang J, Guo J-T. DNA Polymerase alpha is essential for intracellular amplification of hepatitis B virus covalently closed circular DNA. PLoS Pathog 2019; 15:e1007742 [View Article] [PubMed]
     [Google Scholar]
  43. Chisari FV, Mason WS, Seeger C. Virology. Comment on “Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA.”. Science 2014; 344:1237 [View Article] [PubMed]
     [Google Scholar]
  44. Luo J, Cui X, Gao L, Hu J. Identification of an Intermediate in Hepatitis B Virus Covalently Closed Circular (CCC) DNA Formation and Sensitive and Selective CCC DNA Detection. J Virol 2017; 91:e00539-17 [View Article] [PubMed]
     [Google Scholar]
  45. Chin R, Earnest-Silveira L, Koeberlein B, Franz S, Zentgraf H et al. Modulation of MAPK pathways and cell cycle by replicating hepatitis B virus: factors contributing to hepatocarcinogenesis. J Hepatol 2007; 47:325–337 [View Article] [PubMed]
     [Google Scholar]
  46. Wang C-M, Wang Y, Fan C-G, Xu F-F, Sun W-S et al. miR-29c targets TNFAIP3, inhibits cell proliferation and induces apoptosis in hepatitis B virus-related hepatocellular carcinoma. Biochem Biophys Res Commun 2011; 411:586–592 [View Article] [PubMed]
     [Google Scholar]
  47. Lamontagne RJ, Bagga S, Bouchard MJ. Hepatitis B virus molecular biology and pathogenesis. Hepatoma Res 2016; 2:163–186 [View Article] [PubMed]
     [Google Scholar]
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Activities of endogenous APOBEC3s and uracil-DNA-glycosylase affect the hypermutation frequency of hepatitis B virus cccDNA
J. Gen. Virol. 103, 001732 (2022); https://doi.org/10.1099/jgv.0.001732
/content/journal/jgv/10.1099/jgv.0.001732
/content/journal/jgv/10.1099/jgv.0.001732
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