Evolutionary conservation in the hepatitis B virus core structure: comparison of human and duck cores.
BACKGROUND
Hepatitis B virus is a major human pathogen which has been extensively studied, yet its structure is unknown. Cryo-electron microscopy of the viral cores expressed in Escherichia coli or isolated from infected liver provides a means for determining the structure of the hepatitis B nucleocapsid.


RESULTS
Using cryo-electron microscopy and three-dimensional image reconstruction, we have determined the structures of duck and human hepatitis B virus cores and find that they have similar dimer-clustered T = 3 and T = 4 icosahedral organizations. The duck virus core protein sequence differs from the human in both length and amino acid content; however, the only significant structural differences observed are the lobes of density on the lateral edges of the projecting (distal) domain of the core protein dimer. The different cores contain varying amounts of nucleic acid, but exhibit similar contacts between the core protein and the nucleic acid. Immunoelectron microscopy of intact cores has localized two epitopes on the core surface corresponding to residues 76-84 and 129-132.


CONCLUSIONS
The bacterial expression system faithfully reproduces the native hepatitis B virus core structure even in the absence of the complete viral genome. This confirms that proper assembly of the core is independent of genome packaging. Difference imaging and antibody binding map three sequence positions in the structure: the C terminus and the regions near amino acids 80 and 130. Finally, we suggest that the genome-core interactions and the base (proximal) domain of the core dimer are evolutionarily conserved whereas the projecting domain, which interacts with the envelope proteins, is more variable.
Kenney
J M
JM
Structural Biology Programme, EMBL, Heidelberg, Germany.
von Bonsdorff
C H
CH
Nassal
M
M
Fuller
S D
SD
D000818:Animals; D015925:Cryopreservation; D004372:Ducks; D018604:Epitope Mapping; D019143:Evolution, Molecular; D006512:Hepatitis B Core Antigens / Q000276:immunology / Q000648:ultrastructure*; D015611:Hepatitis B Virus, Duck / Q000737:chemistry* / Q000648:ultrastructure; D006515:Hepatitis B virus / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008099:Liver / Q000821:virology; D008854:Microscopy, Electron / Q000379:methods; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D014758:Viral Core Proteins / Q000737:chemistry
D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins; D014758:Viral Core Proteins