AbstractHepadnaviruses are hepatotropic enveloped DNA viruses with an icosahedral capsid. Hepatitis B virus (HBV) causes chronic infection in an estimated 240 million people; Woodchuck Hepatitis virus (WHV), an HBV homologue, has been an important model system for drug development. The dimeric capsid protein (Cp) plays multiple functions during the viral life cycle and thus has become an important target for a new generation of antivirals. Purified HBV and WHV Cp spontaneously assemble into 120-dimer capsids. Though they have 65% identity, WHV Cp has error-prone assembly with stronger protein-protein association. We have taken advantage of the differences in assembly to investigate the basis of assembly regulation. We have determined the structures of the WHV capsid to 4.5 Å resolution by cryo-EM and the WHV Cp dimer to 2.9 Å resolution by crystallography and examined the biophysical properties of the dimer. We found, in dimer, the subdomain that makes protein-protein interactions is partially disordered and rotated 21° from its position in capsid. This subdomain is susceptible to proteolysis, consistent with local disorder. These data show there is an entropic cost for assembly that is compensated for by the energetic gain of burying hydrophobic interprotein contacts. We propose a series of stages in assembly that incorporate disorder-to-order transition and structural shifts. WHV assembly shows similar susceptibility to HBV antiviral molecules, suggesting that HBV assembly follows similar transitions and indicating WHV’s importance as a model system for drug development. We suggest that a cascade of structural changes may be a common mechanism for regulating high fidelity capsid assembly in HBV and other viruses.Author summaryCapsid assembly is a key step in the Hepatitis B virus (HBV) life cycle; it requires sophisticated regulation to ensure the production of capsids and viruses with high-fidelity. HBV capsids are constructed from 120 self-assembling capsid protein dimers (Cp). It was hypothesized that changes in Cp structure regulated the assembly reaction. Here we determined structures of dimer and capsid for Woodchuck Hepatitis virus (WHV), an HBV homologue. We observed that the component of the dimer involved in subunit-subunit interactions undergoes a disorder-order transition and changes structure concomitant with assembly. Meanwhile WHV Cp displays similar susceptibility to HBV antiviral. We propose that this structural transition entropically regulates assembly and may be a common theme in other viruses.
Potential Newer Therapeutic Targets for Hepatitis B Virus Drug Development
