Virologica Sinica

, Volume 33, Issue 2, pp 162–172 | Cite as

Mechanisms and Effects on HBV Replication of the Interaction between HBV Core Protein and Cellular Filamin B

Research Article
  • 109 Downloads

Abstract

Hepatitis B virus (HBV) infection is one of the major problems that threatens global health. There have been many studies on HBV, but the relationship between HBV and host factors is largely unexplored and more studies are needed to clarify these interactions. Filamin B is an actin-binding protein that acts as a cytoskeleton protein, and it is involved in cell development and several signaling pathways. In this study, we showed that filamin B interacted with HBV core protein, and the interaction promoted HBV replication. The interaction between filamin B and core protein was observed in HEK 293T, Huh7 and HepG2 cell lines by co-immunoprecipitation and co-localization immnofluoresence. Overexpression of filamin B increased the levels of HBV total RNAs and pre-genome RNA (pgRNA), and improved the secretion level of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg). In contrast, filamin B knockdown inhibited HBV replication, decreased the level of HBV total RNAs and pgRNA, and reduced the secretion level of HBsAg and HBeAg. In addition, we found that filamin B and core protein may interact with each other via four blocks of argentine residues at the C-terminus of core protein. In conclusion, we identify filamin B as a novel host factor that can interact with core protein to promote HBV replication in hepatocytes. Our study provides new insights into the relationship between HBV and host factors and may provide new strategies for the treatment of HBV infection.

Keywords

Filamin B Core HBV replication 

Notes

Acknowledgements

We thank Prof. Arnoud Sonnenberg (Division of Cell Biology, Netherlands Cancer Institute) for his kind gift of the plasmid Filamin B -GFP. We would also like to thank Prof. Xinwen Chen (State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China) for his kind gift of the plasmid Core -HA. This research was supported by the Postdoctoral Science Foundation of China.

Author Contributions

JFM, DYG and LC conceived the study and participated in the design. YLL and YSS performed the experiments and drafted the manuscript. FYS, RH and CLL analyzed data. JFM and DYG finalized the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflicts of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

This article does not contain any studies with human or animal subjects performed by any of the authors.

Supplementary material

12250_2018_23_MOESM1_ESM.pdf (300 kb)
Supplementary material 1 (PDF 300 kb)

References

  1. Bartenschlager R, Junkerniepmann M, Schaller H (1990) The P-gene product of hepatitis-B virus is required as a structural component for genomic rna encapsidation. J Virol 64:5324–5332PubMedPubMedCentralGoogle Scholar
  2. Beck J, Nassal M (2007) Hepatitis B virus replication. World J Gastroenterol 13:48–64CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bicknell LS, Morgan T, Bonafe L, Wessels MW, Bialer MG, Willems PJ, Cohn DH, Krakow D, Robertson SP (2005) Mutations in FLNB cause boomerang dysplasia. J Med Genet 42:e43CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bicknell LS, Farrington-Rock C, Shafeghati Y, Rump P, Alanay Y, Alembik Y, Al-Madani N, Firth H, Karimi-Nejad MH, Kim CA, Leask K, Maisenbacher M, Moran E, Pappas JG, Prontera P, de Ravel T, Fryns JP, Sweeney E, Fryer A, Unger S, Wilson LC, Lachman RS, Rimoin DL, Cohn DH, Krakow D, Robertson SP (2007) A molecular and clinical study of Larsen syndrome caused by mutations in FLNB. J Med Genet 44:89–98CrossRefPubMedGoogle Scholar
  5. Brocker F, Bardenheuer W, Vieten L, Julicher K, Werner N, Marquitan G, Michael D, Opalka B, Schutte J (1999) Assignment of human filamin gene FLNB to human chromosome band 3p14.3 and identification of YACs containing the complete FLNB transcribed region. Cytogenet Cell Genet 85:267–268CrossRefPubMedGoogle Scholar
  6. Chakarova C, Wehnert MS, Uhl K, Sakthivel S, Vosberg HP, van der Ven PFM, Furst DO (2000) Genomic structure and fine mapping of the two human filamin gene paralogues FLNB and FLNC and comparative analysis of the filamin gene family. Hum Genet 107:597–611CrossRefPubMedGoogle Scholar
  7. Chu CM, Yeh CT, Chien RN, Sheen IS, Liaw YF (1997) The degrees of hepatocyte nuclear but not cytoplasmic expression of hepatitis B core antigen reflect the level of viral replication in chronic hepatitis B virus infection. J Clin Microbiol 35:102–105PubMedPubMedCentralGoogle Scholar
  8. Cooper J, Liu L, Woodruff EA, Taylor HE, Goodwin JS, D’Aquila RT, Spearman P, Hildreth JE, Dong X (2011) Filamin A protein interacts with human immunodeficiency virus type 1 Gag protein and contributes to productive particle assembly. J Biol Chem 286:28498–28510CrossRefPubMedPubMedCentralGoogle Scholar
  9. Cougot D, Neuveut C, Buendia MA (2005) HBV-induced carcinogenesis. J Clin Virol 34:S75–S78CrossRefPubMedGoogle Scholar
  10. del Valle-Perez B, Martinez VG, Lacasa-Salavert C, Figueras A, Shapiro SS, Takafuta T, Casanovas O, Capella G, Ventura F, Vinals F (2010) Filamin B plays a key role in vascular endothelial growth factor-induced endothelial cell motility through its interaction with Rac-1 and Vav-2. J Biol Chem 285:10748–10760CrossRefPubMedPubMedCentralGoogle Scholar
  11. Dotson D, Woodruff EA, Villalta F, Dong XH (2016) Filamin A is involved in HIV-1 Vpu-mediated Evasion of host restriction by modulating tetherin expression. J Biol Chem 291:4236–4246CrossRefPubMedPubMedCentralGoogle Scholar
  12. Dryden KA, Wieland SF, Whitten-Bauer C, Gerin JL, Chisari FV, Yeager M (2006) Native hepatitis B virions and capsids visualized by electron cryomicroscopy. Mol Cell 22:843–850CrossRefPubMedGoogle Scholar
  13. Feng Y, Walsh CA (2004) The many faces of filamin: a versatile molecular scaffold for cell motility and signalling. Nat Cell Biol 6:1034–1038CrossRefPubMedGoogle Scholar
  14. Freed EO (1998) HIV-1 gag proteins: diverse functions in the virus life cycle. Virology 251:1–15CrossRefPubMedGoogle Scholar
  15. Gorlin JB, Yamin R, Egan S, Stewart M, Stossel TP, Kwiatkowski DJ, Hartwig JH (1990) Human endothelial actin-binding protein (Abp-280, Nonmuscle Filamin)—a molecular leaf spring. J Cell Biol 111:1089–1105CrossRefPubMedGoogle Scholar
  16. Guo YH, Li YN, Zhao JR, Zhang J, Yan Z (2011) HBc binds to the CpG islands of HBV cccDNA and promotes an epigenetic permissive state. Epigenetics 6:720–726CrossRefPubMedGoogle Scholar
  17. Hartwig JH, Stossel TP (1975) Isolation and properties of actin, myosin, and a new actinbinding protein in rabbit alveolar macrophages. J Biol Chem 250:5696–5705PubMedGoogle Scholar
  18. Hatton T, Zhou S, Standring DN (1992) RNA- and DNA-binding activities in hepatitis B virus capsid protein: a model for their roles in viral replication. J Virol 66:5232–5241PubMedPubMedCentralGoogle Scholar
  19. Hu J, Lu J, Lian G, Zhang J, Hecht JL, Sheen VL (2014) Filamin B regulates chondrocyte proliferation and differentiation through Cdk1 signaling. PLoS ONE 9:e89352CrossRefPubMedPubMedCentralGoogle Scholar
  20. Huang CJ, Chen YH, Ting LP (2000) Hepatitis B virus core protein interacts with the C-terminal region of actin-binding protein. J Biomed Sci 7:160–168CrossRefPubMedGoogle Scholar
  21. Jeon YJ, Choi JS, Lee JY, Yu KR, Ka SH, Cho Y, Choi EJ, Baek SH, Seol JH, Park D, Bang OS, Chung CH (2008) Filamin B serves as a molecular scaffold for type I interferon-induced c-Jun NH2-terminal kinase signaling pathway. Mol Biol Cell 19:5116–5130CrossRefPubMedPubMedCentralGoogle Scholar
  22. Jia B, Guo M, Li G, Yu D, Zhang X, Lan K, Deng Q (2015) Hepatitis B virus core protein sensitizes hepatocytes to tumor necrosis factor-induced apoptosis by suppression of the phosphorylation of mitogen-activated protein kinase kinase 7. J Virol 89:2041–2051CrossRefPubMedGoogle Scholar
  23. Jimenez-Baranda S, Gomez-Mouton C, Rojas A, Martinez-Prats L, Mira E, Ana Lacalle R, Valencia A, Dimitrov DS, Viola A, Delgado R, Martinez AC, Manes S (2007) Filamin-A regulates actin-dependent clustering of HIV receptors. Nat Cell Biol 9:838–846CrossRefPubMedGoogle Scholar
  24. Jung J, Hwang SG, Chwae YJ, Park S, Shin HJ, Kim K (2014) Phosphoacceptors threonine 162 and serines 170 and 178 within the carboxyl-terminal RRRS/T motif of the hepatitis B virus core protein make multiple contributions to hepatitis B virus replication. J Virol 88:8754–8767CrossRefPubMedPubMedCentralGoogle Scholar
  25. Kann M, Schmitz A, Rabe B (2007) Intracellular transport of hepatitis B virus. World J Gastroenterol 13:39–47CrossRefPubMedPubMedCentralGoogle Scholar
  26. Klumpp K, Shimada T, Allweiss L, Volz T, Lutgehetmann M, Hartman G, Flores OA, Lam AM, Dandri M (2017) Efficacy of NVR 3-778, alone and in combination with pegylated interferon, vs entecavir in uPA/SCID mice with humanized livers and HBV infection. Gastroenterology.  https://doi.org/10.1053/j.gastro.2017.10.017 PubMedGoogle Scholar
  27. Krakow D, Robertson SP, King LM, Morgan T, Sebald ET, Bertolotto C, Wachsmann-Hogiu S, Acuna D, Shapiro SS, Takafuta T, Aftimos S, Kim CA, Firth H, Steiner CE, Cormier-Daire V, Superti-Furga A, Bonafe L, Graham JM Jr, Grix A, Bacino CA, Allanson J, Bialer MG, Lachman RS, Rimoin DL, Cohn DH (2004) Mutations in the gene encoding filamin B disrupt vertebral segmentation, joint formation and skeletogenesis. Nat Genet 36:405–410CrossRefPubMedGoogle Scholar
  28. Lambert C, Doring T, Prange R (2007) Hepatitis B virus maturation is sensitive to functional inhibition of ESCRT-III, vps4, and gamma 2-adaptin. J Virol 81:9050–9060CrossRefPubMedPubMedCentralGoogle Scholar
  29. Liu Q, Rand TA, Kalidas S, Du F, Kim HE, Smith DP, Wang X (2003) R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway. Science 301:1921–1925CrossRefPubMedGoogle Scholar
  30. Liu K, Ludgate L, Yuan Z, Hu J (2015) Regulation of multiple stages of hepadnavirus replication by the carboxyl-terminal domain of viral core protein in trans. J Virol 89:2918–2930CrossRefPubMedGoogle Scholar
  31. Lupberger J, Hildt E (2007) Hepatitis B virus-induced oncogenesis. World J Gastroenterol 13:74–81CrossRefPubMedPubMedCentralGoogle Scholar
  32. Nguyen DH, Ludgate L, Hu J (2008) Hepatitis B virus-cell interactions and pathogenesis. J Cell Physiol 216:289–294CrossRefPubMedPubMedCentralGoogle Scholar
  33. Patrosso MC, Repetto M, Villa A, Milanesi L, Frattini A, Faranda S, Mancini M, Maestrini E, Toniolo D, Vezzoni P (1994) The exon-intron organization of the human X-Linked gene (Fln1) encoding actin-binding protein-280. Genomics 21:71–76CrossRefPubMedGoogle Scholar
  34. Rabe B, Vlachou A, Pante N, Helenius A, Kann M (2003) Nuclear import of hepatitis B virus capsids and release of the viral genome. Proc Natl Acad Sci USA 100:9849–9854CrossRefPubMedPubMedCentralGoogle Scholar
  35. Salfeld J, Pfaff E, Noah M, Schaller H (1989) Antigenic determinants and functional domains in core antigen and e antigen from hepatitis B virus. J Virol 63:798–808PubMedPubMedCentralGoogle Scholar
  36. Scaglioni PP, Melegari M, Wands JR (1997) Posttranscriptional regulation of hepatitis B virus replication by the precore protein. J Virol 71:345–353PubMedPubMedCentralGoogle Scholar
  37. Schadler S, Hildt E (2009) HBV life cycle: entry and morphogenesis. Viruses-Basel 1:185–209CrossRefGoogle Scholar
  38. Serinoz E, Varli M, Erden E, Cinar K, Kansu A, Uzunalimoglu O, Yurdaydin C, Bozkaya H (2003) Nuclear localization of hepatitis B core antigen and its relations to liver injury, hepatocyte proliferation, and viral load. J Clin Gastroenterol 36:269–272CrossRefPubMedGoogle Scholar
  39. Stossel TP, Condeelis J, Cooley L, Hartwig JH, Noegel A, Schleicher M, Shapiro SS (2001) Filamins as integrators of cell mechanics and signalling. Nat Rev Mol Cell Biol 2:138–145CrossRefPubMedGoogle Scholar
  40. Tzeng HT, Tsai HF, Chyuan IT, Liao HJ, Chen CJ, Chen PJ, Hsu PN (2014) Tumor necrosis factor-alpha induced by hepatitis B virus core mediating the immune response for hepatitis B viral clearance in mice model. PLoS ONE 9:e103008CrossRefPubMedPubMedCentralGoogle Scholar
  41. van der Flier A, Sonnenberg A (2001) Structural and functional aspects of filamins. Biochem Biophys Acta 1538:99–117CrossRefPubMedGoogle Scholar
  42. van der Flier A, Kuikman I, Kramer D, Geerts D, Kreft M, Takafuta T, Shapiro SS, Sonnenberg A (2002) Different splice variants of filamin-B affect myogenesis, subcellular distribution, and determine binding to integrin [beta] subunits. J cell Biol 156:361–376CrossRefPubMedPubMedCentralGoogle Scholar
  43. Wang K, Ash JF, Singer SJ (1975) Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells. Proc Natl Acad Sci USA 72:4483–4486CrossRefPubMedPubMedCentralGoogle Scholar
  44. Weigand K, Knaust A, Schaller H (2010) Assembly and export determine the intracellular distribution of hepatitis B virus core protein subunits. J Gen Virol 91:59–67CrossRefPubMedGoogle Scholar
  45. Wills JW, Craven RC (1991) Form, function, and use of retroviral gag proteins. Aids 5:639–654CrossRefPubMedGoogle Scholar
  46. Xie Z, Xu W, Davie EW, Chung DW (1998) Molecular cloning of human ABPL, an actin-binding protein homologue. Biochem Biophys Res Commun 251:914–919CrossRefPubMedGoogle Scholar
  47. Xu WF, Xie ZW, Chung DW, Davie EW (1998) A novel human actin-binding protein homologue that binds to platelet glycoprotein Ib alpha. Blood 92:1268–1276PubMedGoogle Scholar
  48. Yan H, Zhong G, Xu G, He W, Jing Z, Gao Z, Huang Y, Qi Y, Peng B, Wang H, Fu L, Song M, Chen P, Gao W, Ren B, Sun Y, Cai T, Feng X, Sui J, Li W (2012) Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. eLife 1:e00049CrossRefPubMedPubMedCentralGoogle Scholar
  49. Yeh CT, Wong SW, Fung YK, Ou JH (1993) Cell cycle regulation of nuclear localization of hepatitis B virus core protein. Proc Natl Acad Sci USA 90:6459–6463CrossRefPubMedPubMedCentralGoogle Scholar
  50. Yu M, Summers J (1991) A domain of the hepadnavirus capsid protein is specifically required for DNA maturation and virus assembly. J Virol 65:2511–2517PubMedPubMedCentralGoogle Scholar
  51. Zhao K, Wu C, Yao Y, Cao L, Zhang Z, Yuan Y, Wang Y, Pei R, Chen J, Hu X, Zhou Y, Lu M, Chen X (2017) Ceruloplasmin inhibits the production of extracellular hepatitis B virions by targeting its middle surface protein. J Gen Virol 98:1410–1421CrossRefPubMedGoogle Scholar
  52. Zlotnick A, Venkatakrishnan B, Tan Z, Lewellyn E, Turner W, Francis S (2015) Core protein: a pleiotropic keystone in the HBV lifecycle. Antiviral Res 121:82–93CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  1. 1.School of Basic Medical SciencesWuhan UniversityWuhanChina
  2. 2.State Key Laboratory of Virology, Wuhan Institute of VirologyChinese Academy of SciencesWuhanChina
  3. 3.School of Basic Medicine (Shenzhen)Sun Yat-sen UniversityGuangzhouChina

Personalised recommendations