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Journal of Virology, October 2002, p. 10282-10289, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10282-10289.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Promoter Sequences Required for Reactivation of Epstein-Barr Virus from Latency

Ulrich K. Binné,^1, Wolfgang Amon,¹ and Paul J. Farrell^1,2*

Ludwig Institute for Cancer Research,¹ Virology and Cell Biology Section, Imperial College Faculty of Medicine, St. Mary's Campus, London W2 1PG, United Kingdom²

Received 8 April 2002/ Accepted 12 July 2002

A luciferase reporter system with stably transfected oriP plasmids in Akata Burkitt's lymphoma cells provides a quantitative assay for the BZLF1 Zp promoter in response to B-cell receptor (BCR) activation by cross-linking with anti-immunoglobulin. In this system, detailed kinetic studies of promoter activity are possible. Previously reported promoter elements upstream of -221 from the transcription start and the ZIIR sequence had little effect on the Zp promoter, but the ZI and ZIIIA elements were essential for early activation. The ZIIIB element mediates autoactivation. Mutation of the ZV repressor sequence greatly increased the induction of the promoter but did not make it constitutively active. Zp transcription in response to BCR cross-linking declined after a few hours; this decline was reduced and delayed by acyclovir or phosphonoacetic acid, indicating that viral DNA replication or a late viral gene can play a role in the switch off of the Zp promoter. Late expression of the LMP1 protein may account for this.

Present address: Dept. of Molecular Oncology, MGH Cancer Center, Charlestown, MA 02129.

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