Significant interference with hepatitis B virus replication by a core-nuclease fusion protein.
Hepatitis B virus (HBV), a small DNA containing virus that replicates via reverse transcription, causes acute and chronic B-type hepatitis in humans. The limited success of current therapies for chronic infection has prompted exploration of alternative strategies. Capsid-targeted viral inactivation is a conceptually powerful approach that exploits virion structural proteins to target a degradative enzyme specifically into viral particles. Its principal feasibility has been demonstrated in retroviral model systems but not yet for a medically relevant virus outside the retrovirus family. Recently, we found that C proximal fusion to the HBV capsid protein of the Ca(2+)-dependent nuclease (SN) from Staphylococcus aureus yields a chimeric protein, coreSN, that in Escherichia coli coassembles with the wild-type capsid protein into particles with internal SN domains. Here we show that, in HBV co-transfected human hepatoma cells, less than 1 coreSN protein per 10 wild-type core protein subunits reduced titers of enveloped DNA containing virions by more than 95%. The antiviral effect depends on both an enzymatically active SN and on the core domain. CoreSN does not block assembly of RNA containing nucleocapsids but interferes with proper synthesis of viral DNA inside the capsid, or leads to rapid DNA degradation. Our data suggest an intracellular nuclease activation that, owing to the characteristics of HBV morphogenesis, is nonetheless highly virus specific. HBV may therefore be particularly vulnerable to the capsid-targeted viral inactivation approach.
Beterams
G
G
University Hospital Freiburg, Department of Internal Medicine II/Molecular Biology, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Nassal
M
M
D000998:Antiviral Agents / Q000502:physiology*; D002213:Capsid; D004279:DNA, Viral / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D019562:Viral Load; D014779:Virus Replication / Q000502:physiology*
D000998:Antiviral Agents; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins