title: High-resolution methylation analysis and in vivo protein-DNA binding at the promoter of the viral oncogene LMP2A in B cell lines carrying latent Epstein-Barr virus genomes.
abstract: Latency protein LMP2A of Epstein-Barr virus (EBV) has been implicated in EBV related tumorigenesis. To understand the host cell dependent expression of the LMP2A gene, it is necessary to analyse the regulatory mechanisms of the LMP2A promoter (LMP2Ap). By transient transfection and in vitro binding analyses two CBF1 sites have previously been shown to be involved in the regulation of LMP2Ap. However, the promoter structure has not been examined at the nucleotide level in vivo. Therefore we undertook a comprehensive analysis of in vivo protein binding and of CpG-methylation patterns at LMP2Ap in a panel of B cell lines carrying latent EBV genomes. The presence of characteristic footprints on two CBF1 and further binding-sites, together with overall hypomethylation of CpG dinucleotides correlated well with promoter activity. In contrast, the absence of several genomic footprints, as well as the presence of patches of highly methylated CpG dinucleotides were characteristic of silent LMP2Aps.
authors.affiliation: Microbiological Research Group, National Center for Epidemiology, Pihenö ut 1, H-1529 Budapest, Hungary.
authors.forename: Daniel
authors.initials: D
authors.lastname: Salamon
authors.forename: Maria
authors.initials: M
authors.lastname: Takacs
authors.forename: Fritz
authors.initials: F
authors.lastname: Schwarzmann
authors.forename: Hans
authors.initials: H
authors.lastname: Wolf
authors.forename: Janos
authors.initials: J
authors.lastname: Minarovits
authors.forename: Hans Helmut
authors.initials: HH
authors.lastname: Niller
mesh_terms: D001402:B-Lymphocytes / Q000821:virology*; D002460:Cell Line; D002461:Cell Line, Transformed; D018899:CpG Islands / Q000235:genetics; D018983:DNA Footprinting; D019175:DNA Methylation; D016679:Genome, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D011401:Promoter Regions, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D017735:Virus Latency
chemical_list: C037662:EBV-associated membrane antigen, Epstein-Barr virus; D014763:Viral Matrix Proteins