Episomal replication timing of gamma-herpesviruses in latently infected cells.
This study addresses the timing of gammaherpesviral episomal DNA replication with respect to the cell cycle. For the first time we analyzed a rhadinovirus, the prototype Herpesvirus saimiri (HVS), and compared it to the lymphocryptovirus Epstein-Barr virus (EBV). Newly synthesized DNA of latently infected B- or T-cells was first BrdU-labeled; then we sorted the cells corresponding to cell cycle phases G(0/1), G(2/M), and S (4 fractions S(1)-S(4)) and performed anti-BrdU chromatin immunoprecipitation. Next, DNA of different viral gene loci was quantitatively detected together with cellular control genes of known replication time. The sensitive technique is further enhanced by an internal coprecipitation standard for increased precision. Both gammaherpesviruses replicated very early in S-phase, together with cellular euchromatin. Our work suggests that early S-phase DNA replication is a general characteristic of episomal herpesviral genomes.
Vogel
Benjamin
B
Institut für Klinische und Molekulare Virologie, Universitätsklinikum der Friedrich-Alexander-Universität Erlangen-Nürnberg, D-91054 Erlangen, Germany.
Full
Florian
F
Biesinger
Brigitte
B
Linden
Christian
C
Alberter
Barbara
B
Ensser
Armin
A
D001402:B-Lymphocytes / Q000821:virology; D002453:Cell Cycle; D002460:Cell Line; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism; D006567:Herpesvirus 2, Saimiriine / Q000502:physiology*; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D010957:Plasmids; D013601:T-Lymphocytes / Q000821:virology; D013997:Time Factors; D017735:Virus Latency; D014779:Virus Replication
D004279:DNA, Viral