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Fluorescent protein tagging of adenoviral proteins pV and pIX reveals ‘late virion accumulation compartment’
- Source :
- PLoS Pathogens, PLoS Pathogens, Vol 16, Iss 6, p e1008588 (2020)
- Publication Year :
- 2020
- Publisher :
- Public Library of Science, 2020.
-
Abstract
- The human adenovirus type 5 (HAdV5) causes disease of the upper and lower respiratory tract. The early steps of HAdV5 entry up to genome replication in the host nucleus have been extensively studied. However, late stages of infection remain poorly understood. Here, we set out to elucidate the spatiotemporal orchestration of late adenovirus nuclear remodeling in living cells. We generated virus mutants expressing fluorescently tagged protein IX (pIX) and protein V (pV), a capsid and viral genome associated protein, respectively. We found that during progeny virion production both proteins localize to a membrane-less, nuclear compartment, which is highly impermeable such that in immunofluorescence microscopy antibodies can hardly penetrate it. We termed this compartment ‘late virion accumulation compartment’ (LVAC). Correlation between light- and electron microscopy revealed that the LVAC contains paracrystalline arrays of viral capsids that arrange tightly packed within a honeycomb-like organization of viral DNA. Live-cell microscopy as well as FRAP measurements showed that the LVAC is rigid and restricts diffusion of larger molecules, indicating that capsids are trapped inside.<br />Author summary Understanding the regulation of adenovirus morphogenesis is not only of interest to cell biologists but is also key to define novel drug targets as well as to optimize adenoviruses as tools for gene therapy. While early steps of the adenovirus ‘life cycle’ are well understood, it is currently debated how, when and where capsid components associate with viral DNA. Here we used a combination of imaging methods to detail virus-induced spatiotemporal changes at late stages of infection. We found that HAdV5 induces a structured, membrane-less nuclear compartment. In this compartment capsids are closely packed within a honeycomb-like organization of replicated DNA, such that the newly formed particles appear to be trapped and show very little motility. Interestingly, we found a clear discrepancy between immunostaining and fluorescent fusion tagging, indicating a limited penetration of immunostains into this compartment. Since other pathogens induce similar compartments during replication, interpretation of immunostaining data requires careful evaluation.
- Subjects :
- Physiology
viruses
Virus Replication
Biochemistry
Viral Packaging
Virions
Cell Fusion
Adenovirus Infections, Human
Fluorescence Microscopy
Immune Physiology
Fluorescence microscope
Medicine and Health Sciences
Biology (General)
0303 health sciences
Microscopy
Cell fusion
Immune System Proteins
Chemistry
030302 biochemistry & molecular biology
Light Microscopy
Compartment (chemistry)
Lamins
Cell biology
Nucleic acids
Capsid
Research Article
Cell Physiology
QH301-705.5
Immunology
Viral Structure
DNA replication
Research and Analysis Methods
Microbiology
Virus
Antibodies
03 medical and health sciences
Virology
Genetics
Humans
Molecular Biology
030304 developmental biology
Adenoviruses, Human
Virion
Biology and Life Sciences
Proteins
DNA
Cell Biology
RC581-607
Viral Replication
Cytoskeletal Proteins
Viral replication
A549 Cells
DNA, Viral
Parasitology
Capsid Proteins
Immunologic diseases. Allergy
Lamin
Subjects
Details
- Language :
- English
- ISSN :
- 15537374 and 15537366
- Volume :
- 16
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- PLoS Pathogens
- Accession number :
- edsair.doi.dedup.....0d028052c17ca4c78c1c9e597747c2c8