C-1027-induced alterations in Epstein-Barr viral DNA replication in latently infected cultured human Raji cells: relationship to DNA damage.
This study is the first detailing drug-induced changes in EBV DNA replication intermediates (RIs). Both EBV replication inhibition and damage induction were studied in latently infected human Raji cells treated with the enediyne DNA strand-scission agent C-1027. Analysis of RIs on two-dimensional agarose gels revealed a rapid loss in the EBV bubble arc. When elongation of nascent chains was blocked by aphidicolin, this loss was inhibited, suggesting that C-1027-induced disappearance of RIs was related to maturation of preformed replication molecules in the absence of initiation of new RIs. C-1027 damage to EBV DNA was limited at concentrations where loss of the bubble arc was nearly complete, and none was detected within the replicating origin (ori P)-containing fragment, indicating that replication inhibition occurred in trans. By contrast, the non-nuclear mitochondrial genome was insensitive to replication inhibition but highly sensitive to damage induced by C-1027. C-1027-induced trans inhibition of nuclear but not mitochondrial DNA replication is consistent with a cell cycle checkpoint response to a DNA-damaging agent. EBV replication and Raji cell growth were inhibited at equivalent C-1027 doses.
McHugh
M M
MM
Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Beerman
T A
TA
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics / Q000821:virology; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects*; D004272:DNA, Mitochondrial / Q000187:drug effects; D004279:DNA, Viral / Q000187:drug effects*; D015180:Electrophoresis, Gel, Two-Dimensional; D053281:Enediynes; D006131:Growth Inhibitors / Q000494:pharmacology; D004854:Herpesvirus 4, Human / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics / Q000821:virology; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology / Q000821:virology; D017735:Virus Latency / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D004272:DNA, Mitochondrial; D004279:DNA, Viral; D053281:Enediynes; D006131:Growth Inhibitors; C058024:C 1027