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J Virol, April 1998, p. 2969-2974, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Specific Methylation Patterns in Two Control Regions of Epstein-Barr Virus Latency: the LMP-1-Coding Upstream Regulatory Region and an Origin of DNA Replication (oriP)

Kerstin I. Falk,* Laszlo Szekely, Anna Aleman, and Ingemar Ernberg

Microbiology and Tumorbiology Center, Karolinska Institute, S-171 77 Stockholm, Sweden

Received 22 October 1997/Accepted 5 January 1998

The Epstein-Barr virus (EBV) can establish at least four different forms of latent infection. Previously, we have shown that the level of methylation of the EBV genome varies, depending on the form of latency. The methylation status of CpGs was analyzed by the bisulfite genomic sequencing technique in four different cell types representing different forms of latency. The dyad symmetry element of the origin of replication (oriP) region and the latent membrane protein 1 (LMP-1) regulatory sequence (LRS) were studied. The dyad symmetry element has four binding sites for EBNA-1. In a cell with type I latency, a region upstream of the dyad symmetry element was highly methylated, whereas the dyad symmetry element was unmethylated in the EBNA-1-binding region. The LRS was extensively methylated in the LMP-1-negative cell line Rael, in contrast to a LMP-1-expressing nasopharyngeal carcinoma tumor (NPC C15), which was almost completely unmethylated. The methylation pattern of LRS in type I and type III Burkitt lymphoma cells of similar parental origins confirmed that demethylation of some regions takes place upon phenotypic drift.

* Corresponding author. Mailing address: Microbiology and Tumorbiology Center, Karolinska Institute, S-171 77 Stockholm, Sweden. Phone: 46-8-728-6286. Fax: 46-8-319470. E-mail: Kerstin.Falk.{at}mtc.ki.se




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Copyright © 1998 by the American Society for Microbiology. All rights reserved.