Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus.
The 165-kb chromosome of Epstein-Barr virus (EBV) is replicated by cellular enzymes only once per cell cycle in human cells that are latently infected. Here, we report that the human origin recognition complex, ORC, can be detected in association with an EBV replication origin, oriP, in cells by using antibodies against three different subunits of human ORC to precipitate crosslinked chromatin. Mcm2, a subunit of the MCM replication licensing complex, was found to associate with oriP during G(1) and to dissociate from it during S phase. The detection of ORC and Mcm2 at oriP was shown to require the presence of the 120-bp replicator of oriP. Licensing and initiation of replication at oriP of EBV thus seem to be mediated by ORC. This is an example of a virus apparently using ORC and associated factors for the propagation of its genome.
Chaudhuri
B
B
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Xu
H
H
Todorov
I
I
Dutta
A
A
Yates
J L
JL
D002453:Cell Cycle; D002460:Cell Line; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D064111:Minichromosome Maintenance Complex Component 2; D009687:Nuclear Proteins / Q000378:metabolism*; D051738:Origin Recognition Complex; D018741:Replication Origin; D014779:Virus Replication
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C086607:OriP-binding protein 1; D051738:Origin Recognition Complex; D064111:Minichromosome Maintenance Complex Component 2