Abstract
Viruses exploit host factors and environment for their efficient replication. The virus-host interaction mechanisms for achieving an optimal hepatitis B virus (HBV) replication have been largely unknown. Here, a single cell cloning revealed that HepAD38 cells, a widely-used HBV-inducible cell line, contain cell clones with diverse permissiveness to HBV replication. The HBV permissiveness was impaired upon treatment with microtubule inhibitor nocodazole, which was identified as an HBV replication inhibitor from a pharmacological screening. In the microtubule-disrupted cells, the efficiency of HBV capsid assembly was remarkably decreased without significant change in pre-assembly process. We further found that HBV core interacted with tubulin and co-localized with microtubule-like fibriforms, but this association was abrogated upon microtubule-disassembly agents, resulting in attenuation of capsid formation. Our data thus suggest a significant role of microtubules in the efficient capsid formation during HBV replication. In line with this, a highly HBV permissive cell clone of HepAD38 cells showed a prominent association of core-microtubule and thus a high capacity to support the capsid formation. These findings provide a new aspect of virus-cell interaction for rendering efficient HBV replication.
Highlights
Hepatitis B virus (HBV) is a member of the Hepadnaviridae family, a group of enveloped viruses with carrying approximately 3.2 kb relaxed circular DNA as their genome[1, 2]
hepatitis B virus (HBV) virions produced from Hep38.7-Tet cells showed similar infectivity to that from the parental HepAD38 cells, examined in the HepaRG cell infection assay with viral inoculum being normalizing by HBV DNA copy genome equivalent (Fig. 1B)
These steps are reported to be regulated by host factors including nuclear factor 1 (HNF-1), CCAAT binding factor (CBF), Rab[7], Vps[4] and the endosomal sorting complex required for transport (ESCRT) III29–33
Summary
Hepatitis B virus (HBV) is a member of the Hepadnaviridae family, a group of enveloped viruses with carrying approximately 3.2 kb relaxed circular DNA (rcDNA) as their genome[1, 2]. HBV genome encodes four major open reading frames for core, polymerase, surface, and x proteins. Core and polymerase are especially essential for viral DNA replication. One of the transcripts with approximately 3.5 kb in length, called pre-genomic (pg) RNA, plays an essential role in HBV replication. PgRNA encodes viral polymerase and core proteins. While polymerase interacts with pgRNA, core proteins spontaneously dimerize and multimerize to assemble into the capsids. Screening of a pharmacological inhibitor library using a highly HBV-permissive cell clone revealed that microtubules played a significant role in supporting the process for HBV capsid assembly. We investigated a relevance of the core-microtubule association in the host permissiveness to HBV replication
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