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Hepatitis B virus virion secretion is a CRM1-spike-mediated late event
BACKGROUND: Hepatitis B virus (HBV) is a major human pathogen worldwide. To date, there is no curative treatment for chronic hepatitis B. The mechanism of virion secretion remains to be investigated. Previously, we found that nuclear export of HBc particles can be facilitated via two CRM1-specific n...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9210616/ https://www.ncbi.nlm.nih.gov/pubmed/35729569 http://dx.doi.org/10.1186/s12929-022-00827-w |
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author | Su, Pei-Yi Yen, Shin-Chwen Bruce Yang, Ching-Chun Chang, Chih-Hsu Lin, Wen-Chang Shih, Chiaho |
author_facet | Su, Pei-Yi Yen, Shin-Chwen Bruce Yang, Ching-Chun Chang, Chih-Hsu Lin, Wen-Chang Shih, Chiaho |
author_sort | Su, Pei-Yi |
collection | PubMed |
description | BACKGROUND: Hepatitis B virus (HBV) is a major human pathogen worldwide. To date, there is no curative treatment for chronic hepatitis B. The mechanism of virion secretion remains to be investigated. Previously, we found that nuclear export of HBc particles can be facilitated via two CRM1-specific nuclear export signals (NES) at the spike tip. METHODS: In this study, we used site-directed mutagenesis at the CRM1 NES, as well as treatment with CRM1 inhibitors at a low concentration, or CRM1-specific shRNA knockdown, in HBV-producing cell culture, and measured the secretion of various HBV viral and subviral particles via a native agarose gel electrophoresis assay. Separated HBV particles were characterized by Western blot analysis, and their genomic DNA contents were measured by Southern blot analysis. Secreted extracellular particles were compared with intracellular HBc capsids for DNA synthesis and capsid formation. Virion secretion and the in vivo interactions among HBc capsids, CRM1 and microtubules, were examined by proximity ligation assay, immunofluorescence microscopy, and nocodazole treatment. RESULTS: We report here that the tip of spike of HBV core (HBc) particles (capsids) contains a complex sensor for secretion of both HBV virions and naked capsids. HBV virion secretion is closely associated with HBc nuclear export in a CRM1-dependent manner. At the conformationally flexible spike tips of HBc particles, NES motifs overlap extensively with motifs important for secretion of HBV virions and naked capsids. CONCLUSIONS: We provided experimental evidence that virions and naked capsids can egress via two distinct, yet overlapping, pathways. Unlike the secretion of naked capsids, HBV virion secretion is highly CRM1- and microtubule-dependent. CRM1 is well known for its involvement in nuclear transport in literature. To our knowledge, this is the first report that CRM1 is required for virion secretion. CRM1 inhibitors could be a promising therapeutic candidate for chronic HBV patients in clinical medicine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12929-022-00827-w. |
format | Online Article Text |
id | pubmed-9210616 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-92106162022-06-22 Hepatitis B virus virion secretion is a CRM1-spike-mediated late event Su, Pei-Yi Yen, Shin-Chwen Bruce Yang, Ching-Chun Chang, Chih-Hsu Lin, Wen-Chang Shih, Chiaho J Biomed Sci Research BACKGROUND: Hepatitis B virus (HBV) is a major human pathogen worldwide. To date, there is no curative treatment for chronic hepatitis B. The mechanism of virion secretion remains to be investigated. Previously, we found that nuclear export of HBc particles can be facilitated via two CRM1-specific nuclear export signals (NES) at the spike tip. METHODS: In this study, we used site-directed mutagenesis at the CRM1 NES, as well as treatment with CRM1 inhibitors at a low concentration, or CRM1-specific shRNA knockdown, in HBV-producing cell culture, and measured the secretion of various HBV viral and subviral particles via a native agarose gel electrophoresis assay. Separated HBV particles were characterized by Western blot analysis, and their genomic DNA contents were measured by Southern blot analysis. Secreted extracellular particles were compared with intracellular HBc capsids for DNA synthesis and capsid formation. Virion secretion and the in vivo interactions among HBc capsids, CRM1 and microtubules, were examined by proximity ligation assay, immunofluorescence microscopy, and nocodazole treatment. RESULTS: We report here that the tip of spike of HBV core (HBc) particles (capsids) contains a complex sensor for secretion of both HBV virions and naked capsids. HBV virion secretion is closely associated with HBc nuclear export in a CRM1-dependent manner. At the conformationally flexible spike tips of HBc particles, NES motifs overlap extensively with motifs important for secretion of HBV virions and naked capsids. CONCLUSIONS: We provided experimental evidence that virions and naked capsids can egress via two distinct, yet overlapping, pathways. Unlike the secretion of naked capsids, HBV virion secretion is highly CRM1- and microtubule-dependent. CRM1 is well known for its involvement in nuclear transport in literature. To our knowledge, this is the first report that CRM1 is required for virion secretion. CRM1 inhibitors could be a promising therapeutic candidate for chronic HBV patients in clinical medicine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12929-022-00827-w. BioMed Central 2022-06-21 /pmc/articles/PMC9210616/ /pubmed/35729569 http://dx.doi.org/10.1186/s12929-022-00827-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Su, Pei-Yi Yen, Shin-Chwen Bruce Yang, Ching-Chun Chang, Chih-Hsu Lin, Wen-Chang Shih, Chiaho Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title | Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title_full | Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title_fullStr | Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title_full_unstemmed | Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title_short | Hepatitis B virus virion secretion is a CRM1-spike-mediated late event |
title_sort | hepatitis b virus virion secretion is a crm1-spike-mediated late event |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9210616/ https://www.ncbi.nlm.nih.gov/pubmed/35729569 http://dx.doi.org/10.1186/s12929-022-00827-w |
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