Interaction between hepatitis B virus core protein and reverse transcriptase.
Previous mutagenesis studies with hepatitis B virus (HBV) suggest that continued interactions with core are required for several steps in genomic replication. To examine core-polymerase (Pol) interactions, insect cells were coinfected with baculovirus constructs that independently expressed core and Pol. The results demonstrated several features with implications that core plays an interactive role with HBV Pol: (i) core coprecipitated with constructs expressing full-length Pol as well as the terminal protein (TP), reverse transcriptase (RT) and RNase H domains of Pol, independently; (ii) coprecipitation of core was not dependent on the presence of an epsilon stem-loop sequence; and (iii) core-Pol complexes migrated as intact capsid particles, as detected by sucrose gradient analysis. To analyze the structural and sequence requirements of core in recognition of Pol, a series of core mutants with two- to four-amino-acid insertions or carboxy-terminal deletions were assessed for Pol interaction. The results indicated that capsid formation is required but not sufficient for interaction with Pol and that the TP and RT domains of Pol have different requirements for interaction with core. To map the core binding sites on Pol, a panel of amino- and carboxy-terminal deletion mutants of the TP and RT domains of Pol were analyzed for interaction with core. At least three separate core binding sites on Pol were detected. This analysis begins to define basic requirements for core-Pol interactions, but further study is necessary to delineate the effects of these interactions on encapsidation and genome replication.
Lott
L
L
Department of Virology and Immunology, Southwest Regional Primate Research Center, Southwest Foundation for Biomedical Research, San Antonio, Texas 78227, USA.
Beames
B
B
Notvall
L
L
Lanford
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D006515:Hepatitis B virus / Q000502:physiology*; D008969:Molecular Sequence Data; D011485:Protein Binding; D012194:RNA-Directed DNA Polymerase / Q000502:physiology*; D014758:Viral Core Proteins / Q000502:physiology*; D014779:Virus Replication
D014758:Viral Core Proteins; D012194:RNA-Directed DNA Polymerase