Formation of intranuclear replication compartments of Epstein-Barr virus with redistribution of BZLF1 and BMRF1 gene products.

The localizations of the Epstein-Barr virus immediate-early transcriptional activator BZLF1 protein ZEBRA, of the BMRF1 early antigen diffuse component (EA-D), and of viral DNA replication were studied in the Burkitt's lymphoma cell line Akata treated with anti-human immunoglobulin antibodies. Prompt and sequential appearance of ZEBRA, EA-D, and viral DNA was observed in about 70% of the cells. At early times after activation, ZEBRA had a diffuse intranuclear distribution, but later it was concentrated in globular regions within the nucleus. EA-D appeared first in a finely stippled pattern and then in a diffuse pattern. At late times, EA-D concentrated in globular regions similar to those with ZEBRA. Double staining for ZEBRA and EA-D revealed that ZEBRA followed the morphological changes of EA-D with a 1-2 hr delay and that both finally coalesced in the same structures, where in situ hybridization localized replicating viral DNA. The redistribution of both ZEBRA and EA-D to these compartments depended upon the replication of lytic viral DNA. These findings indicate that these globular regions are sites for viral replication and that transcription of EBV late genes may be regulated in these structures.