Relating structure to function in the hepatitis delta virus antigen.
Hepatitis delta virus expresses two forms of a single protein, the small (delta Ag-S) and large (delta Ag-L) antigens, which are identical except for an additional 19 residues present at the C terminus of delta Ag-L. While delta Ag-S is required to promote genome replication, delta Ag-L potently inhibits this process and also facilitates packaging of the viral genome by envelope proteins of the helper virus (hepatitis B virus). Regions within the antigens responsible for nuclear localization, RNA binding, and dimerization have been identified, yet it is not clear how these particular activities contribute to the ultimate replication and packaging phenotypes. Here we report the following findings. (i) Although the removal of the nuclear localization signal from either antigen resulted in significant cytoplasmic accumulation, both proteins still had access to the nucleus. As a consequence, no functional defect was observed with either mutant. (ii) The RNA-binding domain, although necessary for delta Ag-S function, could be deleted from delta Ag-L without compromising its ability to either inhibit replication or promote packaging. (iii) In contrast, the coiled-coil dimerization domain was required for both the activation of replication by delta Ag-S and the inhibition of replication by delta Ag-L. This region, with an additional 20 amino acids C-terminal to it, was necessary and sufficient to potently inhibit replication by interacting with the small antigen. (iv) The packaging property of delta Ag-L required a C-terminal Pro/Gly-rich region which is hypothesized to interact with the hepatitis B virus envelope proteins during the assembly process.
Lazinski
D W
DW
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111.
Taylor
J M
JM
D000956:Antigens, Viral / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002451:Cell Compartmentation; D002467:Cell Nucleus; D004252:DNA Mutational Analysis; D005455:Fluorescent Antibody Technique; D003698:Hepatitis Delta Virus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D035923:Hepatitis delta Antigens; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D021382:Protein Sorting Signals; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D013329:Structure-Activity Relationship; D014779:Virus Replication
D000956:Antigens, Viral; D035923:Hepatitis delta Antigens; D021382:Protein Sorting Signals; D016601:RNA-Binding Proteins; C442244:hepatitis delta virus large antigen