Interaction between hepatitis B virus core protein and reverse transcriptase

L. Lott, B. Beames, L. Notvall, R. E. Lanford

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

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 and that the TP and RT domains of 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 were analyzed for interaction with core. At least three separate core binding sites on 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.

Original languageEnglish (US)
Pages (from-to)11479-11489
Number of pages11
JournalJournal of Virology
Volume74
Issue number24
DOIs
StatePublished - 2000
Externally publishedYes

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Viral Core Proteins
Hepatitis B virus
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
capsid
Capsid
binding sites
Binding Sites
Inverted Repeat Sequences
Ribonuclease H
coprecipitation
mutants
proteins
Baculoviridae
Mutant Proteins
mutagenesis
Cell Communication
Mutagenesis
Sequence Analysis
Insects

ASJC Scopus subject areas

  • Immunology

Cite this

Interaction between hepatitis B virus core protein and reverse transcriptase. / Lott, L.; Beames, B.; Notvall, L.; Lanford, R. E.

In: Journal of Virology, Vol. 74, No. 24, 2000, p. 11479-11489.

Research output: Contribution to journalArticle

Lott, L. ; Beames, B. ; Notvall, L. ; Lanford, R. E. / Interaction between hepatitis B virus core protein and reverse transcriptase. In: Journal of Virology. 2000 ; Vol. 74, No. 24. pp. 11479-11489.
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