PMID: 9525596Apr 3, 1998Paper

Generation of replication-competent hepatitis B virus nucleocapsids in insect cells

Journal of Virology
M SeiferD N Standring

Abstract

The double-stranded DNA genome of human hepatitis B virus (HBV) and related hepadnaviruses is reverse transcribed from a pregenomic RNA by a viral polymerase (Pol) harboring both priming and RNA- and DNA-dependent elongation activities. Although hepadnavirus replication occurs inside viral nucleocapsids, or cores, biochemical systems for analyzing this reaction are currently limited to unencapsidated Pols expressed in heterologous systems. Here, we describe cis and trans classes of replicative HBV cores, produced in the recombinant baculovirus system via coexpression of HBV core and Pol proteins from either a single RNA (i.e., in cis) or two distinct RNAs (in trans). Upon isolation from insect cells, cis and trans cores contained Pol-linked HBV minus-strand DNA with 5' ends mapping to the authentic elongation origin DR1 and also plus-strand DNA species. Only trans cores, however, were highly active for the de novo priming and reverse transcription of authentic HBV minus strands in in vitro endogenous polymerase assays. This reaction strictly required HBV Pol but not the epsilon stem-loop element, although the presence of one epsilon, or better, two epsilons, enhanced minus-strand synthesis up to 10-fold. Compared to unencapsida...Continue Reading

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Citations

Apr 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·S UrbanD L Tyrrell
Nov 26, 1998·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·X XiongC S Gibbs
Feb 7, 2002·Expert Opinion on Investigational Drugs·William DelaneyStephen A Locarnini
Feb 27, 2008·World Journal of Gastroenterology : WJG·Lin-Lin GaoYan Li
Jun 29, 2007·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Anna S LokUNKNOWN Hepatitis B Virus Drug Resistance Working Group
Dec 1, 2005·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Boerries BrandenburgEberhard Hildt
Dec 3, 1998·Antimicrobial Agents and Chemotherapy·M SeiferD N Standring
Jan 8, 2004·Journal of Hepatology·Stephen Locarnini

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