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 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.
Microbiologie Fondamentale et Pathogénicité, MFP CNRS UMR 5234, Université de Bordeaux, Bordeaux 33076, France; Department of Infection Biology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
Tetsuro
T
Komatsu
Microbiologie Fondamentale et Pathogénicité, MFP CNRS UMR 5234, Université de Bordeaux, Bordeaux 33076, France; Bordeaux INP, MCMP, UMR 5234, Bordeaux 33000, France.
Denis
D
Dacheux
Department of Gene Therapy, Ulm University, Ulm 89081, Germany.
Florian
F
Kreppel
Department of Infection Biology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
Kyosuke
K
Nagata
Microbiologie Fondamentale et Pathogénicité, MFP CNRS UMR 5234, Université de Bordeaux, Bordeaux 33076, France.
Harald
H
Wodrich
D000256:Adenoviridae / Q000378:metabolism*; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D006801:Humans
D002843:Chromatin