Expression of hepatitis B virus core antigen gene in Saccharomyces cerevisiae: synthesis of two polypeptides translated from different initiation codons.
Two recombinant plasmids were constructed that allow expression of the hepatitis B core (HBc) antigen gene in the yeast Saccharomyces cerevisiae under the control of the repressible acid phosphatase promoter. One plasmid was designed to produce polypeptide I, which consists of 183 amino acids, and the other plasmid was designed to produce polypeptide II, which has an additional 29-amino-acid sequence at the amino terminus of polypeptide I. The viral genome may code for either one or both of these two polypeptides, depending upon the selection of initiation codons. Both polypeptides produced in yeast cells reacted with anti-HBc antibody and were assembled into spherical particles approximately 27 nm in diameter. Particles made of polypeptide I were stable, whereas those made of polypeptide II readily dissociated when exposed to high salt levels. The antigenicity of the HBc (as defined by its reactivity to anti-HBc antibody in the reversed passive hemagglutination assay) disappeared as the particle dissociated, leaving materials that sedimented slowly and that reacted to anti-hepatitis B e antibody. These observations strongly suggest that native viral cores are mostly (if not all) made of polypeptide I, because it is reasonably stable, and that the N-terminal portion of this polypeptide has some, but not a profound, influence on the assembly of polypeptides into particles.
D002213:Capsid / Q000235:genetics; D003001:Cloning, Molecular; D003062:Codon; D005786:Gene Expression Regulation; D005814:Genes, Viral; D006510:Hepatitis B Antibodies / Q000276:immunology; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D008854:Microscopy, Electron; D008970:Molecular Weight; D009024:Morphogenesis; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014758:Viral Core Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D003062:Codon; D006510:Hepatitis B Antibodies; D006512:Hepatitis B Core Antigens; D014758:Viral Core Proteins; D014764:Viral Proteins