The Epstein-Barr virus major latent promoter Qp is constitutively active, hypomethylated, and methylation sensitive

Qian Tao, Keith D Robertson, Angela Manns, Allan Hildesheim, Richard F. Ambinder

Research output: Contribution to journalArticle

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Abstract

Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is indispensable for viral DNA replication and episome maintenance in latency. Four promoters, Cp, Wp, Qp, and Fp, are known to drive EBNA1 expression. Here we show that the TATA-less Qp is constitutively active in a variety of EBV-positive [EBV(+)] tumors and cell lines, irrespective of the activities of other EBNA1 promoters, the type of viral latency, and the cell type. The transcription of highly regulated promoters such as the EBV Cp is known to be directly regulated by CpG methylation. To characterize the role of CpG methylation in the regulation of the constitutively active Qp, we performed bisulfite genomic sequencing and functional analyses using a methylation cassette transcriptional reporter assay. Twenty consecutive CpG sites (16 proximal to the Qp initiation site and 4 upstream of the adjacent Fp initiation site) were studied by bisulfite sequencing of DNA extracted from EBV(+) tumors and cell lines. Eighteen EBV(+) tumors of lymphoid (B, T, and NK cell) or epithelial origin and five Burkitt's lymphoma cell lines were studied. The 16 CpG sites proximal to Qp were virtually all unmethylated, but the 4 CpG sites upstream of the Fp initiation site were variably methylated. The methylation cassette assay showed that in vitro methylation of the Qp cassette (-172 to +32) resulted in strong repression of Qp activity in transient transfection. Thus, Qp is susceptible to repression by methylation but was found to be consistently hypomethylated and expressed in all tumors and tumor-derived cell lines studied.

Original languageEnglish (US)
Pages (from-to)7075-7083
Number of pages9
JournalJournal of Virology
Volume72
Issue number9
StatePublished - Sep 1998
Externally publishedYes

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Human herpesvirus 4
Human Herpesvirus 4
methylation
Methylation
promoter regions
nuclear antigens
Epstein-Barr Virus Nuclear Antigens
Tumor Cell Line
neoplasms
cell lines
bisulfites
Virus Latency
Burkitt Lymphoma
Viral DNA
natural killer cells
assays
DNA replication
transfection
lymphoma
DNA Replication

ASJC Scopus subject areas

  • Immunology

Cite this

The Epstein-Barr virus major latent promoter Qp is constitutively active, hypomethylated, and methylation sensitive. / Tao, Qian; Robertson, Keith D; Manns, Angela; Hildesheim, Allan; Ambinder, Richard F.

In: Journal of Virology, Vol. 72, No. 9, 09.1998, p. 7075-7083.

Research output: Contribution to journalArticle

Tao, Qian ; Robertson, Keith D ; Manns, Angela ; Hildesheim, Allan ; Ambinder, Richard F. / The Epstein-Barr virus major latent promoter Qp is constitutively active, hypomethylated, and methylation sensitive. In: Journal of Virology. 1998 ; Vol. 72, No. 9. pp. 7075-7083.
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