Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.

Through evolution, Hepatitis B Virus (HBV) developed highly intricate mechanisms exploiting host resources for its multiplication within a constrained genetic coding capacity. Yet a clear picture of viral hitchhiking of cellular processes with spatial resolution is still largely unsolved. Here, by l...

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Autores principales:	Lei Yue, Chang Li, Mingzhu Xu, Min Wu, Jiahui Ding, Jiangxia Liu, Xiaonan Zhang, Zhenghong Yuan
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/7f8c17db42b549cc87485dbc0353c6a7
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spelling	oai:doaj.org-article:7f8c17db42b549cc87485dbc0353c6a72021-12-02T20:00:17ZProbing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.1553-73661553-737410.1371/journal.ppat.1009838https://doaj.org/article/7f8c17db42b549cc87485dbc0353c6a72021-08-01T00:00:00Zhttps://doi.org/10.1371/journal.ppat.1009838https://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Through evolution, Hepatitis B Virus (HBV) developed highly intricate mechanisms exploiting host resources for its multiplication within a constrained genetic coding capacity. Yet a clear picture of viral hitchhiking of cellular processes with spatial resolution is still largely unsolved. Here, by leveraging bDNA-based fluorescence in situ hybridization (FISH) combined with immunofluorescence, we developed a microscopic approach for multiplex detection of viral nucleic acids and proteins, which enabled us to probe some of the key aspects of HBV life cycle. We confirmed the slow kinetics and revealed the high variability of viral replication at single-cell level. We directly visualized HBV minichromosome in contact with acetylated histone 3 and RNA polymerase II and observed HBV-induced degradation of Smc5/6 complex only in primary hepatocytes. We quantified the frequency of HBV pregenomic RNAs occupied by translating ribosome or capsids. Statistics at molecular level suggested a rapid translation phase followed by a slow encapsidation and maturation phase. Finally, the roles of microtubules (MTs) on nucleocapsid assembly and virion morphogenesis were analyzed. Disruption of MTs resulted in the perinuclear retention of nucleocapsid. Meanwhile, large multivesicular body (MVB) formation was significantly disturbed as evidenced by the increase in number and decrease in volume of CD63+ vesicles, thus inhibiting mature virion secretion. In conclusion, these data provided spatially resolved molecular snapshots in the context of specific subcellular activities. The heterogeneity observed at single-cell level afforded valuable molecular insights which are otherwise unavailable from bulk measurements.Lei YueChang LiMingzhu XuMin WuJiahui DingJiangxia LiuXiaonan ZhangZhenghong YuanPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 17, Iss 8, p e1009838 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Lei Yue Chang Li Mingzhu Xu Min Wu Jiahui Ding Jiangxia Liu Xiaonan Zhang Zhenghong Yuan Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
description	Through evolution, Hepatitis B Virus (HBV) developed highly intricate mechanisms exploiting host resources for its multiplication within a constrained genetic coding capacity. Yet a clear picture of viral hitchhiking of cellular processes with spatial resolution is still largely unsolved. Here, by leveraging bDNA-based fluorescence in situ hybridization (FISH) combined with immunofluorescence, we developed a microscopic approach for multiplex detection of viral nucleic acids and proteins, which enabled us to probe some of the key aspects of HBV life cycle. We confirmed the slow kinetics and revealed the high variability of viral replication at single-cell level. We directly visualized HBV minichromosome in contact with acetylated histone 3 and RNA polymerase II and observed HBV-induced degradation of Smc5/6 complex only in primary hepatocytes. We quantified the frequency of HBV pregenomic RNAs occupied by translating ribosome or capsids. Statistics at molecular level suggested a rapid translation phase followed by a slow encapsidation and maturation phase. Finally, the roles of microtubules (MTs) on nucleocapsid assembly and virion morphogenesis were analyzed. Disruption of MTs resulted in the perinuclear retention of nucleocapsid. Meanwhile, large multivesicular body (MVB) formation was significantly disturbed as evidenced by the increase in number and decrease in volume of CD63+ vesicles, thus inhibiting mature virion secretion. In conclusion, these data provided spatially resolved molecular snapshots in the context of specific subcellular activities. The heterogeneity observed at single-cell level afforded valuable molecular insights which are otherwise unavailable from bulk measurements.
format	article
author	Lei Yue Chang Li Mingzhu Xu Min Wu Jiahui Ding Jiangxia Liu Xiaonan Zhang Zhenghong Yuan
author_facet	Lei Yue Chang Li Mingzhu Xu Min Wu Jiahui Ding Jiangxia Liu Xiaonan Zhang Zhenghong Yuan
author_sort	Lei Yue
title	Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
title_short	Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
title_full	Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
title_fullStr	Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
title_full_unstemmed	Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.
title_sort	probing the spatiotemporal patterns of hbv multiplication reveals novel features of its subcellular processes.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/7f8c17db42b549cc87485dbc0353c6a7
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Probing the spatiotemporal patterns of HBV multiplication reveals novel features of its subcellular processes.

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