Generation of Covalently Closed Circular DNA of Hepatitis B Viruses via Intracellular Recycling Is Regulated in a Virus Specific Manner

English version České info

Persistence of hepatitis B virus (HBV) infection requires covalently closed circular (ccc)DNA formation and amplification, which can occur via intracellular recycling of the viral polymerase-linked relaxed circular (rc) DNA genomes present in virions. Here we reveal a fundamental difference between HBV and the related duck hepatitis B virus (DHBV) in the recycling mechanism. Direct comparison of HBV and DHBV cccDNA amplification in cross-species transfection experiments showed that, in the same human cell background, DHBV but not HBV rcDNA converts efficiently into cccDNA. By characterizing the distinct forms of HBV and DHBV rcDNA accumulating in the cells we find that nuclear import, complete versus partial release from the capsid and complete versus partial removal of the covalently bound polymerase contribute to limiting HBV cccDNA formation; particularly, we identify genome region-selectively opened nuclear capsids as a putative novel HBV uncoating intermediate. However, the presence in the nucleus of around 40% of completely uncoated rcDNA that lacks most if not all of the covalently bound protein strongly suggests a major block further downstream that operates in the HBV but not DHBV recycling pathway. In summary, our results uncover an unexpected contribution of the virus to cccDNA formation that might help to better understand the persistence of HBV infection. Moreover, efficient DHBV cccDNA formation in human hepatoma cells should greatly facilitate experimental identification, and possibly inhibition, of the human cell factors involved in the process.

Vyšlo v časopise: Generation of Covalently Closed Circular DNA of Hepatitis B Viruses via Intracellular Recycling Is Regulated in a Virus Specific Manner. PLoS Pathog 6(9): e32767. doi:10.1371/journal.ppat.1001082
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001082

Souhrn

Zdroje

1. ChuCM

LiawYF

2009 Incidence and risk factors of progression to cirrhosis in inactive carriers of hepatitis B virus. Am J Gastroenterol 104 1693 1699

2. KaoJH

ChenDS

2002 Global control of hepatitis B virus infection. Lancet Infect Dis 2 395 403

3. BeckJ

NassalM

2007 Hepatitis B virus replication. World J Gastroenterol 13 48 64

4. NassalM

2008 Hepatitis B viruses: reverse transcription a different way. Virus Res 134 235 249

5. GerlichWH

RobinsonWS

1980 Hepatitis B virus contains protein attached to the 5′ terminus of its complete DNA strand. Cell 21 801 809

6. LocarniniS

MasonWS

2006 Cellular and virological mechanisms of HBV drug resistance. J Hepatol 44 422 431

7. TuttlemanJS

PourcelC

SummersJ

1986 Formation of the pool of covalently closed circular viral DNA in hepadnavirus-infected cells. Cell 47 451 460

8. NassalM

2009 New insights into HBV replication: new opportunities for improved therapies. Future Virology 4 55 70

9. Werle-LapostolleB

BowdenS

LocarniniS

WursthornK

PetersenJ

2004 Persistence of cccDNA during the natural history of chronic hepatitis B and decline during adefovir dipivoxil therapy. Gastroenterology 126 1750 1758

10. BourneEJ

DienstagJL

LopezVA

SanderTJ

LongletJM

2007 Quantitative analysis of HBV cccDNA from clinical specimens: correlation with clinical and virological response during antiviral therapy. J Viral Hepat 14 55 63

11. WielandSF

SpangenbergHC

ThimmeR

PurcellRH

ChisariFV

2004 Expansion and contraction of the hepatitis B virus transcriptional template in infected chimpanzees. Proc Natl Acad Sci U S A 101 2129 2134

12. GaoW

HuJ

2007 Formation of hepatitis B virus covalently closed circular DNA: removal of genome-linked protein. J Virol 81 6164 6174

13. GuoH

JiangD

ZhouT

CuconatiA

BlockTM

2007 Characterization of the intracellular deproteinized relaxed circular DNA of hepatitis B virus: an intermediate of covalently closed circular DNA formation. J Virol 81 12472 12484

14. SunD

NassalM

2006 Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus. J Hepatol 45 636 645

15. ChisariFV

1996 Hepatitis B virus transgenic mice: models of viral immunobiology and pathogenesis. Curr Top Microbiol Immunol 206 149 173

16. RaneyAK

EggersCM

KlineEF

GuidottiLG

PontoglioM

2001 Nuclear covalently closed circular viral genomic DNA in the liver of hepatocyte nuclear factor 1 alpha-null hepatitis B virus transgenic mice. J Virol 75 2900 2911

17. SchultzU

GrgacicE

NassalM

2004 Duck hepatitis B virus: an invaluable model system for HBV infection. Adv Virus Res 63 1 70

18. DallmeierK

SchultzU

NassalM

2008 Heterologous replacement of the supposed host determining region of avihepadnaviruses: high in vivo infectivity despite low infectivity for hepatocytes. PLoS Pathog 4 e1000230

19. ZhangYY

ZhangBH

TheeleD

LitwinS

TollE

2003 Single-cell analysis of covalently closed circular DNA copy numbers in a hepadnavirus-infected liver. Proc Natl Acad Sci U S A 100 12372 12377

20. SummersJ

SmithPM

HorwichAL

1990 Hepadnavirus envelope proteins regulate covalently closed circular DNA amplification. J Virol 64 2819 2824

21. SummersJ

SmithPM

HuangMJ

YuMS

1991 Morphogenetic and regulatory effects of mutations in the envelope proteins of an avian hepadnavirus. J Virol 65 1310 1317

22. MillerRH

RobinsonWS

1984 Hepatitis B virus DNA forms in nuclear and cytoplasmic fractions of infected human liver. Virology 137 390 399

23. LevreroM

PollicinoT

PetersenJ

BelloniL

RaimondoG

2009 Control of cccDNA function in hepatitis B virus infection. J Hepatol 51 581 592

24. KannM

SchmitzA

RabeB

2007 Intracellular transport of hepatitis B virus. World J Gastroenterol 13 39 47

25. RabeB

DelaleauM

BischofA

FossM

SominskayaI

2009 Nuclear entry of hepatitis B virus capsids involves disintegration to protein dimers followed by nuclear reassociation to capsids. PLoS Pathog 5 e1000563

26. GuoH

MaoR

BlockTM

GuoJT

2010 Production and function of the cytoplasmic deproteinized relaxed circular DNA of hepadnaviruses. J Virol 84 387 396

27. KöckJ

BlumHE

2008 Hypermutation of hepatitis B virus genomes by APOBEC3G, APOBEC3C and APOBEC3H. J Gen Virol 89 1184 1191

28. PughJC

YaginumaK

KoikeK

SummersJ

1988 Duck hepatitis B virus (DHBV) particles produced by transient expression of DHBV DNA in a human hepatoma cell line are infectious in vitro. J Virol 62 3513 3516

29. MarzluffWFJr

1978 Transcription of RNA in isolated nuclei. Methods Cell Biol 19 317 332

30. KöckJ

BaumertTF

DelaneyWEt

BlumHE

von WeizsäckerF

2003 Inhibitory effect of adefovir and lamivudine on the initiation of hepatitis B virus infection in primary tupaia hepatocytes. Hepatology 38 1410 1418

31. CaoF

TavisJE

2004 Detection and characterization of cytoplasmic hepatitis B virus reverse transcriptase. J Gen Virol 85 3353 3360

32. AbrahamTM

LewellynEB

HainesKM

LoebDD

2008 Characterization of the contribution of spliced RNAs of hepatitis B virus to DNA synthesis in transfected cultures of Huh7 and HepG2 cells. Virology 379 30 37

33. KöckJ

NassalM

DeresK

BlumHE

von WeizsäckerF

2004 Hepatitis B virus nucleocapsids formed by carboxy-terminally mutated core proteins contain spliced viral genomes but lack full-size DNA. J Virol 78 13812 13818

34. AddisonWR

WaltersKA

WongWW

WilsonJS

MadejD

2002 Half-life of the duck hepatitis B virus covalently closed circular DNA pool in vivo following inhibition of viral replication. J Virol 76 6356 6363

35. ZhuY

YamamotoT

CullenJ

SaputelliJ

AldrichCE

2001 Kinetics of hepadnavirus loss from the liver during inhibition of viral DNA synthesis. J Virol 75 311 322

36. GuoJT

PryceM

WangX

BarrasaMI

HuJ

2003 Conditional replication of duck hepatitis B virus in hepatoma cells. J Virol 77 1885 1893

37. ZhouT

GuoH

GuoJT

CuconatiA

MehtaA

2006 Hepatitis B virus e antigen production is dependent upon covalently closed circular (ccc) DNA in HepAD38 cell cultures and may serve as a cccDNA surrogate in antiviral screening assays. Antiviral Res 72 116 124

38. SällbergM

RudenU

MagniusLO

HarthusHP

NoahM

1991 Characterisation of a linear binding site for a monoclonal antibody to hepatitis B core antigen. J Med Virol 33 248 252

39. StevenAC

ConwayJF

ChengN

WattsNR

BelnapDM

2005 Structure, assembly, and antigenicity of hepatitis B virus capsid proteins. Adv Virus Res 64 125 164

40. LeeGH

WasserS

LimSG

2008 Hepatitis B pregenomic RNA splicing–the products, the regulatory mechanisms and its biological significance. Virus Res 136 1 7

41. ConnellyJC

LeachDR

2004 Repair of DNA covalently linked to protein. Mol Cell 13 307 316

42. YangW

SummersJ

1998 Infection of ducklings with virus particles containing linear double-stranded duck hepatitis B virus DNA: illegitimate replication and reversion. J Virol 72 8710 8717

43. HantzO

ParentR

DurantelD

GriponP

Guguen-GuillouzoC

2009 Persistence of the hepatitis B virus covalently closed circular DNA in HepaRG human hepatocyte-like cells. J Gen Virol 90 127 135

44. GriponP

RuminS

UrbanS

Le SeyecJ

GlaiseD

2002 Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci U S A 99 15655 15660

45. DallmeierK

NassalM

2008 Hepadnaviruses have a narrow host range - do they?

WeberO

ProtzerU

Comparative Hepatitis Basel, Boston Birkhäuser

46. NassalM

LeiferI

WingertI

DallmeierK

PrinzS

2007 A structural model for duck hepatitis B virus core protein derived by extensive mutagenesis. J Virol 81 13218 13229

47. BasagoudanavarSH

PerlmanDH

HuJ

2007 Regulation of hepadnavirus reverse transcription by dynamic nucleocapsid phosphorylation. J Virol 81 1641 1649

48. MelegariM

WolfSK

SchneiderRJ

2005 Hepatitis B virus DNA replication is coordinated by core protein serine phosphorylation and HBx expression. J Virol 79 9810 9820

49. BouchardMJ

SchneiderRJ

2004 The enigmatic X gene of hepatitis B virus. J Virol 78 12725 12734

50. ChenHS

KanekoS

GironesR

AndersonRW

HornbuckleWE

1993 The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol 67 1218 1226

51. ZoulimF

SaputelliJ

SeegerC

1994 Woodchuck hepatitis virus X protein is required for viral infection in vivo. J Virol 68 2026 2030

52. JilbertAR

MillerDS

ScougallCA

TurnbullH

BurrellCJ

1996 Kinetics of duck hepatitis B virus infection following low dose virus inoculation: one virus DNA genome is infectious in neonatal ducks. Virology 226 338 345

53. PasekM

GotoT

GilbertW

ZinkB

SchallerH

1979 Hepatitis B virus genes and their expression in E. coli. Nature 282 575 579

54. MandartE

KayA

GalibertF

1984 Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences. J Virol 49 782 792

55. NassalM

1992 The arginine-rich domain of the hepatitis B virus core protein is required for pregenome encapsidation and productive viral positive-strand DNA synthesis but not for virus assembly. J Virol 66 4107 4116

56. LenhoffRJ

SummersJ

1994 Coordinate regulation of replication and virus assembly by the large envelope protein of an avian hepadnavirus. J Virol 68 4565 4571

57. BeckJ

VogelM

NassalM

2002 dNTP versus NTP discrimination by phenylalanine 451 in duck hepatitis B virus P protein indicates a common structure of the dNTP-binding pocket with other reverse transcriptases. Nucleic Acids Res 30 1679 1687

58. VorreiterJ

LeiferI

RöslerC

JackevicaL

PumpensP

2007 Monoclonal antibodies providing topological information on the duck hepatitis B virus core protein and avihepadnaviral nucleocapsid structure. J Virol 81 13230 13234