dc.contributor.author |
Lennemann, Nicholas J |
|
dc.contributor.author |
Evans, Azia S |
|
dc.contributor.author |
Coyne, Carolyn B |
|
dc.date.accessioned |
2021-04-16T19:51:28Z |
|
dc.date.available |
2021-04-16T19:51:28Z |
|
dc.date.issued |
2020-09-25 |
|
dc.identifier |
v12101074 |
|
dc.identifier.issn |
1999-4915 |
|
dc.identifier.issn |
1999-4915 |
|
dc.identifier.uri |
https://hdl.handle.net/10161/22574 |
|
dc.description.abstract |
Enteroviruses manipulate host membranes to form replication organelles, which concentrate
viral and host factors to allow for efficient replication. However, this process has
not been well-studied in living cells throughout the course of infection. To define
the dynamic process of enterovirus membrane remodeling of major secretory pathway
organelles, we have developed plasmid-based reporter systems that utilize viral protease-dependent
release of a nuclear-localized fluorescent protein from the endoplasmic reticulum
(ER) membrane during infection, while retaining organelle-specific fluorescent protein
markers such as the ER and Golgi. This system thus allows for the monitoring of organelle-specific
changes induced by infection in real-time. Using long-term time-lapse imaging of living
cells infected with coxsackievirus B3 (CVB), we detected reporter translocation to
the nucleus beginning ~4 h post-infection, which correlated with a loss of Golgi integrity
and a collapse of the peripheral ER. Lastly, we applied our system to study the effects
of a calcium channel inhibitor, 2APB, on virus-induced manipulation of host membranes.
We found that 2APB treatment had no effect on the kinetics of infection or the percentage
of infected cells. However, we observed aberrant ER structures in CVB-infected cells
treated with 2APB and a significant decrease in viral-dependent cell lysis, which
corresponded with a decrease in extracellular virus titers. Thus, our system provides
a tractable platform to monitor the effects of inhibitors, gene silencing, and/or
gene editing on viral manipulation of host membranes, which can help determine the
mechanism of action for antivirals.
|
|
dc.language |
eng |
|
dc.publisher |
MDPI AG |
|
dc.relation.ispartof |
Viruses |
|
dc.relation.isversionof |
10.3390/v12101074 |
|
dc.subject |
Cell Line, Tumor |
|
dc.subject |
Intracellular Membranes |
|
dc.subject |
Endoplasmic Reticulum |
|
dc.subject |
Golgi Apparatus |
|
dc.subject |
Humans |
|
dc.subject |
Enterovirus B, Human |
|
dc.subject |
Calcium Channel Blockers |
|
dc.subject |
Virus Replication |
|
dc.subject |
Kinetics |
|
dc.subject |
Genes, Reporter |
|
dc.subject |
Plasmids |
|
dc.subject |
Host-Pathogen Interactions |
|
dc.subject |
Secretory Pathway |
|
dc.subject |
Optical Imaging |
|
dc.title |
Imaging-Based Reporter Systems to Define CVB-Induced Membrane Remodeling in Living
Cells.
|
|
dc.type |
Journal article |
|
duke.contributor.id |
Coyne, Carolyn B|1086848 |
|
dc.date.updated |
2021-04-16T19:51:26Z |
|
pubs.begin-page |
1074 |
|
pubs.end-page |
1074 |
|
pubs.issue |
10 |
|
pubs.organisational-group |
School of Medicine |
|
pubs.organisational-group |
Molecular Genetics and Microbiology |
|
pubs.organisational-group |
Duke |
|
pubs.organisational-group |
Basic Science Departments |
|
pubs.publication-status |
Published |
|
pubs.volume |
12 |
|
duke.contributor.orcid |
Coyne, Carolyn B|0000-0002-1884-6309 |
|