Stability governs the apparent expression of "particulate" hepatitis B e antigen by mutant hepatitis B virus core particles.
The p21.5 capsid or core protein of hepatitis B virus carries two distinct classes of epitopes. Core (HBc) epitopes are found exclusively on the surface of the 28-nm viral icosahedral capsids or core particles, while HBe epitopes are normally expressed only by subparticulate forms of the core protein. Recent studies have suggested that a "particulate" form of HBe is expressed on the surface of capsid particles assembled from p17, a truncated core protein that lacks the carboxy-terminal protamine-like region of p21.5 and hence the ability to bind and encapsidate RNA. In this report we have used epitope-specific ELISAs in conjunction with capsids assembled from a series of carboxy-terminally truncated core proteins to address the mechanistic basis for particulate HBe. Specifically, we sought to test the idea that particulate HBe expression might be linked to the loss of RNA binding. However, our results strongly suggest that expression of HBe by mutant core particles is a result of their intrinsic instability which increases sharply when RNA binding is lost. We show that core particles assembled from mutant core proteins lacking Cys residues also express HBe, again because of capsid instability. We report mild conditions that can induce the dissociation of the mutant capsids and discuss our findings in terms of the factors that control capsid stability.
Seifer
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Hormone Research Institute, University of California, San Francisco 94143-0534.
Standring
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli; D005615:Freezing; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000096:biosynthesis* / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D014981:Xenopus
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens; D012367:RNA, Viral; D011994:Recombinant Proteins; D003545:Cysteine