Live Cell Imaging Reveals HBV Capsid Translocation from the Nucleus To the Cytoplasm Enabled by Cell Division

Romero, Sofia; Unchwaniwala, Nuruddin; Evans, Edward L.; Eliceiri, Kevin W.; Loeb, Daniel D.; Sherer, Nathan M.

doi:10.1128/mbio.03303-22

Cited by 3 publications

(5 citation statements)

References 82 publications

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“…The absence of free core detection in the cell environment is consistent with the rapid kinetic of capsid assembly as previously reported [77]. Of note, the intracellular distribution of capsids would not depend on time of harvest since all experiments were performed at 72 h. post-transfection [42]. LHBs proteins were seen to oligomerize forming a crescent shape at the nuclear periphery as well as large dots dispersed within the cytoplasm (Fig.…”

Section: Discussionsupporting

confidence: 87%

“…Recently, we, among others, documented the intracellular co-localization of LHBs and core proteins in HBV-permissive cells and concluded that LHBs would serve as a platform for HBV virion assembly by recruiting core and SHBs proteins [35,41,42]. Here we used a FRET assay to visualize in cell the core-LHBs interaction when these two partners are co-expressed using a set of tagged proteins: core-eGFP, eGFP-LHBs or mCherry-LHBs as a tracer (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…By TEM, wt cores are detected as capsids with a regular electron-dense shape (Fig. 4) [36,42] in contrast with Y132A-core that does not form capsids (Fig. 3A and Fig.…”

Section: Discussionmentioning

confidence: 99%

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The Hepatitis B virus large envelope protein initiates virion assembly by recruiting pre-assembled capsids at membrane rich domains related to late endosome

Seigneuret,

Eymieux,

Sarabia-Vega

et al. 2024

Preprint

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A crucial step of HBV (Hepatitis B Virus) virion morphogenesis is the envelopment of the nucleocapsid by the viral envelope proteins, which is triggered by an interaction between the HBV core protein and the large HBV envelope protein. To document this interaction, we co-expressed core and large HBV envelope (LHBs) in Huh-7 cells and subjected the cells to microscopy examination by Fluorescence Resonance Energy Transfer (FRET) and Transmission Electron Microscopy (TEM). Our results show that the sole expression of the core protein leads to assembly of capsids that remain individually isolated within the whole cell, but particularly within the nucleus. In the presence of LHBs, capsids were observed as large clusters in a membrane rich region peripheral to the nucleus. In this context, core-LHBs complex co-localize with markers of the late endosome/multivesicular bodies, this co-localization being driven by LHBs. These results thus show that LHBs binds to the core proteins when preassembled into capsid, at membranes of the late endosome, where the inner capsid and the outer envelope meet to assemble a virion.

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Section: Discussionsupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

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The Hepatitis B virus large envelope protein initiates virion assembly by recruiting pre-assembled capsids at membrane rich domains related to late endosome

Seigneuret,

Eymieux,

Sarabia-Vega

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Next, we asked whether the plasmid form of the viral genome was sufficient to induce replication stress. We transiently transfected HepG2 cells with the HBV-encoding plasmid TMA153 [hereafter referred to as pHBV; (58)] and performed DFA 24 hours post-transfection (Fig. 3E-H).…”

Section: Hbv-induced Replication Stress Increases As Infection Progre...mentioning

confidence: 99%

“…The media on transfected cells was replaced with fresh prewarmed medium between 6 to 18 hours post-transfection. The TMA153 plasmid referred to in this study as pHBV, is the same as the previously published pTMA153 and pLJ144, expressing full length HBV pgRNA, Cp, P, and all 3 envelope proteins (58). The pHBVHBeAg plasmid, previously referred to as pHC-9/3142, possesses a deletion in the cis-acting signal for pgRNA encapsidation, leading to the lack of production of the E antigen.…”

Section: Plasmids and Transfectionsmentioning

confidence: 99%

Hepatitis B Virus genomes associate with cellular sites of DNA damage by inducing replication stress

Marcoe,

Larsen,

Majumder

2024

Preprint

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Hepatitis B Virus (HBV) is a leading cause of liver cancer, with almost 300 million infected individuals worldwide. Although HBV-infected patients benefit from drug regimens that help to control chronic infection, they are rarely clinically cured of HBV. The HBV genome persists in the nucleus of infected hepatocytes in the form of a covalently closed circular DNA (cccDNA) molecule, a reservoir of HBV DNA molecules that serve as the template for reactivation of long-term chronic HBV. However, despite playing a central role in the viral life cycle, little is understood about where cccDNA molecules localize, why they are so stable, and how they impact the host nuclear compartment. Perhaps because of this, there are few treatments that target cccDNA, which is critical for eradication of clinical HBV. Here, we show that HBV infection induces a cellular DNA Damage Response (DDR) that is comparable with cells undergoing replication stress, and this cellular replication stress is initiatedafterthe formation of cellular cccDNA molecules. Using a novel high-throughput chromosome conformation capture technology that monitors the localization of HBV cccDNA molecules, we show that cccDNA molecules persist in the vicinity of many cellular fragile sites. Induction of cellular DNA damage leads to relocalization of the viral HBx oncoprotein to DDR sites in an ATM, ATR and DNA-PK dependent manner. Our findings contribute to the understanding of how HBV cccDNA navigates the host nuclear environment, identifying functional targets for development of therapies against HBV infection and resulting liver cancer.ImportanceHepatitis B Virus (HBV) is the leading infectious cause of liver cancer globally. The virus persists in the nucleus long term by forming reservoirs in human liver cells. We have discovered that the HBV DNA localizes to sites on the host genome associated with DNA damage, and in doing so, HBV interferes with the host’s ability to efficiently amplify itself. This results in the induction of cellular DNA breaks, which we propose contributes to eventual cancer progression. Our findings provide new insights into how HBV infection may lead to liver cancer.

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Retroviral hijacking of host transport pathways for genome nuclear export

Behrens,

Sherer

2023

mBio

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Recent advances in the study of virus-cell interactions have improved our understanding of how viruses that replicate their genomes in the nucleus (e.g., retroviruses, hepadnaviruses, herpesviruses, and a subset of RNA viruses) hijack cellular pathways to export these genomes to the cytoplasm where they access virion egress pathways. These findings shed light on novel aspects of viral life cycles relevant to the development of new antiviral strategies and can yield new tractable, virus-based tools for exposing additional secrets of the cell. The goal of this review is to summarize defined and emerging modes of virus-host interactions that drive the transit of viral genomes out of the nucleus across the nuclear envelope barrier, with an emphasis on retroviruses that are most extensively studied. In this context, we prioritize discussion of recent progress in understanding the trafficking and function of the human immunodeficiency virus type 1 Rev protein, exemplifying a relatively refined example of stepwise, cooperativity-driven viral subversion of multi-subunit host transport receptor complexes.

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Live Cell Imaging Reveals HBV Capsid Translocation from the Nucleus To the Cytoplasm Enabled by Cell Division

Abstract: Hepatitis B virus (HBV) is an enveloped, reverse-transcribing DNA virus that is a major cause of liver disease and hepatocellular carcinoma. Subcellular trafficking events underpinning HBV capsid assembly and virion egress remain poorly characterized.

Cited by 3 publications

References 82 publications

The Hepatitis B virus large envelope protein initiates virion assembly by recruiting pre-assembled capsids at membrane rich domains related to late endosome

The Hepatitis B virus large envelope protein initiates virion assembly by recruiting pre-assembled capsids at membrane rich domains related to late endosome

Hepatitis B Virus genomes associate with cellular sites of DNA damage by inducing replication stress

Retroviral hijacking of host transport pathways for genome nuclear export

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