A hydrophobic heptad repeat of the core protein of woodchuck hepatitis virus is required for capsid assembly.
The capsid particle of hepadnaviruses is assembled from its dimer precursors. However, the mechanism of the protein-protein interaction is still poorly understood. A small region in the capsid protein of woodchuck hepatitis virus (WHV) contains four hydrophobic residues, including leucine 101, leucine 108, valine 115, and phenylalanine 122, that are conserved and spaced every seventh residue in the primary sequence to form a hydrophobic heptad repeat (hhr). A hydrophobic force often plays an important role in the interaction of proteins. Therefore, to investigate the role of this region in capsid assembly, we individually changed the codons specifying these four hydrophobic amino acids to codons specifying alanine or proline. In addition, we examined the in vivo infectivity of a WHV genome bearing a naturally occurring single amino acid change (histidine 104-->proline) in the hhr region. The phenotype of each altered genome was determined in both eukaryotic and prokaryotic systems by a capsid protein assay and electron microscopic examination. We show that replacement of any one of the four hydrophobic residues with alanine did not prevent capsid assembly. However, assembled capsid particles were not detected if combinations of any two of the four residues were substituted with alanines or if the spacing of these four hydrophobic residues was changed. An individual introduction of a proline (which dramatically changes the secondary structure of proteins) into different positions of this small region also abolished capsid assembly in vitro or viral replication in vivo. These results suggested that the hhr region of the core protein of WHV was critical for capsid assembly.
Yu
M
M
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892-0470, USA.
Miller
R H
RH
Emerson
S
S
Purcell
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000378:metabolism*; D017919:Hepatitis B Virus, Woodchuck / Q000502:physiology*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D012091:Repetitive Sequences, Nucleic Acid; D016415:Sequence Alignment; D017421:Sequence Analysis; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D019065:Virus Assembly / Q000235:genetics*
D014758:Viral Core Proteins