PMID- 34370796
OWN - NLM
STAT- MEDLINE
DCOM- 20211129
LR  - 20211129
IS  - 1553-7374 (Electronic)
IS  - 1553-7366 (Print)
IS  - 1553-7366 (Linking)
VI  - 17
IP  - 8
DP  - 2021 Aug
TI  - Probing the spatiotemporal patterns of HBV multiplication reveals novel features 
      of its subcellular processes.
PG  - e1009838
LID - 10.1371/journal.ppat.1009838 [doi]
LID - e1009838
AB  - 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.
FAU - Yue, Lei
AU  - Yue L
AD  - Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic 
      Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
FAU - Li, Chang
AU  - Li C
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
FAU - Xu, Mingzhu
AU  - Xu M
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
FAU - Wu, Min
AU  - Wu M
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
FAU - Ding, Jiahui
AU  - Ding J
AD  - Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic 
      Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.
FAU - Liu, Jiangxia
AU  - Liu J
AD  - Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic 
      Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.
FAU - Zhang, Xiaonan
AU  - Zhang X
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
AD  - Centre for Research in Therapeutic Solutions, Biomedical Sciences, Faculty of 
      Science and Technology, University of Canberra, ACT, Australia.
FAU - Yuan, Zhenghong
AU  - Yuan Z
AUID- ORCID: 0000-0003-0268-4891
AD  - Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic 
      Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.
AD  - Research Unit, Shanghai Public Health Clinical Center, Fudan University, 
      Shanghai, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210809
PL  - United States
TA  - PLoS Pathog
JT  - PLoS pathogens
JID - 101238921
RN  - 0 (DNA, Viral)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - DNA, Viral/*analysis
MH  - Hep G2 Cells
MH  - Hepatitis B/*virology
MH  - Hepatitis B virus/*physiology
MH  - Humans
MH  - RNA, Viral/*analysis
MH  - Spatio-Temporal Analysis
MH  - Subcellular Fractions/*virology
MH  - Virion
MH  - *Virus Assembly
MH  - *Virus Replication
PMC - PMC8376071
COIS- The authors declared no competing interests exists.
EDAT- 2021/08/10 06:00
MHDA- 2021/11/30 06:00
PMCR- 2021/08/09
CRDT- 2021/08/09 17:22
PHST- 2021/02/16 00:00 [received]
PHST- 2021/07/25 00:00 [accepted]
PHST- 2021/08/19 00:00 [revised]
PHST- 2021/08/10 06:00 [pubmed]
PHST- 2021/11/30 06:00 [medline]
PHST- 2021/08/09 17:22 [entrez]
PHST- 2021/08/09 00:00 [pmc-release]
AID - PPATHOGENS-D-21-00354 [pii]
AID - 10.1371/journal.ppat.1009838 [doi]
PST - epublish
SO  - PLoS Pathog. 2021 Aug 9;17(8):e1009838. doi: 10.1371/journal.ppat.1009838. 
      eCollection 2021 Aug.