A system for studying the selective encapsidation of hepadnavirus RNA

Abstract

All hepadnaviruses produce multiple genome-length RNA species, only one of which is encapsidated into subviral core particles prior to reverse transcription. To study the encapsidation mechanism, we developed a system in which the packaging of genetically marked target genomes of duck hepatitis B virus is mediated by factors supplied from a separate (helper) plasmid that encodes encapsidation functions. In the helper plasmid, the synthesis of the viral core and polymerase proteins was driven by a simian virus 40 promoter; the RNA produced by this construct was itself inefficiently packaged and was not active as a template for reverse transcription. Cotransfection of this construct with mutant genomes bearing frameshift lesions in either core or polymerase cistrons resulted in the successful packaging and reverse transcription of the mutant genomes. This system should allow definition of both the cis- and trans-acting elements of the encapsidation pathway.