A Method for Visualization of Incoming Adenovirus Chromatin Complexes in Fixed and Living Cells

Inside the adenovirus virion, the genome forms a chromatin-like structure with viral basic core proteins. Core protein VII is the major DNA binding protein and was shown to remain associated with viral genomes upon virus entry even after nuclear delivery. It has been suggested that protein VII plays...

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Bibliographic Details
Published in:PloS one 2015-09, Vol.10 (9), p.e0137102-e0137102
Main Authors: Komatsu, Tetsuro, Dacheux, Denis, Kreppel, Florian, Nagata, Kyosuke, Wodrich, Harald
Format: Article
Language:English
Subjects:
Adenoviridae - metabolism
Adenoviruses
Cell Line
Cells (biology)
Chromatin
Chromatin - metabolism
Core protein
Deoxyribonucleic acid
DNA
Gene expression
Genetic aspects
Genomes
Health aspects
Humans
Immunoglobulins
In vivo methods and tests
Infections
Labeling
Life Sciences
Methods
Microbiology and Parasitology
Monoclonal antibodies
Nuclear transport
Physiological aspects
Plasmids
Polypeptides
Proteins
Virions
Virology
Viruses
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Summary:Inside the adenovirus virion, the genome forms a chromatin-like structure with viral basic core proteins. Core protein VII is the major DNA binding protein and was shown to remain associated with viral genomes upon virus entry even after nuclear delivery. It has been suggested that protein VII plays a regulatory role in viral gene expression and is a functional component of viral chromatin complexes in host cells. As such, protein VII could be used as a maker to track adenoviral chromatin complexes in vivo. In this study, we characterize a new monoclonal antibody against protein VII that stains incoming viral chromatin complexes following nuclear import. Furthermore, we describe the development of a novel imaging system that uses Template Activating Factor-I (TAF-I/SET), a cellular chromatin protein tightly bound to protein VII upon infection. This setup allows us not only to rapidly visualize protein VII foci in fixed cells but also to monitor their movement in living cells. These powerful tools can provide novel insights into the spatio-temporal regulation of incoming adenoviral chromatin complexes.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0137102