A tryptophan-rich motif in the carboxyl terminus of the small envelope protein of hepatitis B virus is central to the assembly of hepatitis delta virus particles.
The small hepatitis B virus surface antigen (S-HBsAg) is capable of driving the assembly and secretion of hepatitis delta virus (HDV) particles by interacting with the HDV ribonucleoprotein (RNP). Previously, a specific domain of the S-HBsAg protein carboxyl terminus, including a tryptophan residue at position 196 (W196), was proven essential for HDV maturation (S. Jenna and C. Sureau, J. Virol. 73: 3351-3358, 1999). Mutation of W196 to phenylalanine (W196F) was permissive for HBV subviral particle (SVP) secretion but deleterious to HDV virion assembly. Here, the W196F S-HBsAg deficiency was assigned to a loss of its ability for interaction with the large HDV antigen (L-HDAg), a major component of the RNP. Because the overall S-HBsAg carboxyl terminus is particularly rich in tryptophan, an amino acid frequently involved in protein-protein interactions, site-directed mutagenesis was conducted to investigate the function of the S-HBsAg Trp-rich domain in HDV assembly. Single substitutions of tryptophan between positions 163 and 201 with alanine or phenylalanine were tolerated for SVP secretion, but those affecting W196, W199, and W201 were detrimental for HDV assembly. This was proven to result from a reduced capacity of the mutants for interaction with L-HDAg. In addition, a W196S S-HBsAg mutant, which has been described in HBV strains that arose in a few cases of lamivudine-treated HBV-infected patients, was deficient for HDV assembly as a consequence of its impaired capacity for interacting with L-HDAg. Interestingly, the fact that even the most conservative substitution of phenylalanine for tryptophan at positions 196, 199, or 201 was sufficient to ablate interaction of S-HBsAg with L-HDAg suggests that W196, W199, and W201 are located at a binding interface that is central to HDV maturation.
Komla-Soukha
Isabelle
I
Laboratoire de Virologie, Institut National de la Transfusion Sanguine, 6 rue Alexandre Cabanel, 75739, Paris, France.
Sureau
Camille
C
D020816:Amino Acid Motifs / Q000502:physiology; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D045744:Cell Line, Tumor; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D003698:Hepatitis Delta Virus / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism* / Q000502:physiology; D011392:Proline / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism* / Q000502:physiology; D014771:Virion / Q000378:metabolism* / Q000502:physiology; D019065:Virus Assembly / Q000502:physiology*
D006514:Hepatitis B Surface Antigens; D010446:Peptide Fragments; C118669:S envelope protein, hepatitis B virus; D014759:Viral Envelope Proteins; D014364:Tryptophan; D011392:Proline