The duck hepatitis B virus P-gene codes for protein strongly associated with the 5'-end of the viral DNA minus strand

Abstract

A number of antisera, elicited against different segments of the duck hepatitis B virus (DHBV) P-gene translation product, were used to immunoprecipitate the protein that is covalently bound to the 5'-end of the DHBV DNA minus strand. For monitoring purposes, a small DNA minus-strand fragment, carrying this protein, was radioactively labeled. All of the P-specific antisera specifically immunoprecipitated this DNA fragment demonstrating that the protein species attached to the immunoprecipitated DNA fragment were products of the DHBV P-gene. The electrophoretic behavior, in SDS gels, of the DNA minus-strand fragment-protein complex indicated that it was present mostly in the form of aggregates. However, a small fraction consisted of DNA minus-strand fragments carrying P-gene proteins, encoded solely within the 5'-region of the P-gene. This indicated that different P-gene proteins, presumably covalently bound at a common region and subsequently processed, were bound to the 5'-end of the DHBV DNA minus strand. The DHBV P-gene presumably codes for the virus-associated reverse transcriptase and DNA polymerase activities. Using the P-gene-specific antisera, it was not possible to detect putative P-gene-coded polymerase proteins in a free form, i.e., not bound to viral DNA. This may be due to insufficient sensitivity or to the polymerase protein(s) being heterogeneous and/or aggregated. In addition, it is possible that the genome-bound protein itself may have polymerase activity.