In vivo phosphorylation and protein analysis of hepatitis B virus core antigen.
The C open reading frame of the hepatitis B virus contains two in-frame ATG codons that are separated by the precore region and encodes two major polypeptides that are antigenically distinct and that are probably synthesized from individual mRNAs. The precore region directs the secretion of the e antigen, whereas the core antigen can be expressed in the absence of these sequences. In this report a transient expression system was used to study the hepatitis B virus core antigen. By using a chimeric complex of adenovirus major late promoter-simian virus 40 enhancer sequences, we were able to achieve high levels of core antigen expression in transfected cells, permitting characterization of this protein and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The core polypeptide is a 20.9-kilodalton protein, and we show in this study that it is phosphorylated in vivo. Cell fractionation studies, the results of which are supported by indirect immunofluorescence, localized the phosphocore in the cytosol and the nucleus and indicated that it is associated with the membrane of transfected cells. Results of Triton X-114 solubilization studies indicated that the phosphocore is peripherally associated with cytoplasmic membranes. Expression of the membrane-associated phosphocore occurred in the absence of the precore sequences. The phosphocore also assembled into particles in the absence of other viral gene products or intact DNA.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006512:Hepatitis B Core Antigens / Q000235:genetics* / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification; D010766:Phosphorylation; D010957:Plasmids
D000596:Amino Acids; D006512:Hepatitis B Core Antigens; D008565:Membrane Proteins