Methylated DNA Recognition during the Reversal of Epigenetic Silencing Is Regulated by Cysteine and Serine Residues in the Epstein-Barr Virus Lytic Switch Protein
Open accessAbstract
Epstein-Barr virus (EBV) causes infectious mononucleosis and is associated with various malignancies, including Burkitt's lymphoma and nasopharyngeal carcinoma. Like all herpesviruses, the EBV life cycle alternates between latency and lytic replication. During latency, the viral genome is largely silenced by host-driven methylation of CpG motifs and, in the switch to the lytic cycle, this epigenetic silencing is overturned. A key event is the activation of the viral BRLF1 gene by the immediate-early protein Zta. Zta is a bZIP transcription factor that preferentially binds to specific response elements (ZREs) in the BRLF1 promoter (Rp) when these elements are methylated. Zta's ability to trigger lytic cycle activation is severely compromised when a cysteine residue in its bZIP domain is mutated to serine (C189S), but the molecular basis for this effect is unknown. Here we show that the C189S mutant is defective for activating Rp in a Burkitt's lymphoma cell line. The mutant is compromised both in vitro and in vivo for binding two methylated ZREs in Rp (ZRE2 and ZRE3), although the effect is striking only for ZRE3. Molecular modeling of Zta bound to methylated ZRE3, together with biochemical data, indicate that C189 directly contacts one of the two methyl cytosines within a specific CpG motif. The motif's second methyl cytosine (on the complementary DNA strand) is predicted to contact S186, a residue known to regulate methyl-ZRE recognition. Our results suggest that C189 regulates the enhanced interaction of Zta with methylated DNA in overturning the epigenetic control of viral latency. As C189 is conserved in many bZIP proteins, the selectivity of Zta for methylated DNA may be a paradigm for a more general phenomenon.
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One cysteine and one serine amino acid interact with 5-methylcytosine.
“Two amino acids, one cysteine and one serine, were found to interact with 5mC45.”
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- Authors
- Karlsson, Q. H., Schelcher, C., Verrall, E., Petosa, C., & Sinclair, A. J.
- Affiliations
- University of Sussex (GB)
- European Molecular Biology Laboratory (FR)
- Corresponding author
- Sinclair, Alison J.
- Venue
- PLoS Pathogens
- Publisher
- Public Library of Science (PLoS)
- Published
- 2008-03-07
- Volume
- 4
- Issue
- 3
- Pages
- e1000005
- ISSN
- 1553-7366 (ISSN-L)
- 1553-7374
- PMC
- PMC2267006
- OpenAlex
- W1967863146
- Cited by
- 39
- References
- 26
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 2 · 2021: 2 · 2018: 1 · 2017: 2 · 2016: 4 · 2015: 2 · 2013: 4 · 2012: 4
- FWCI
- 2.19
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS Pathogens (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Sussex Research Online (University of Sussex) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Research Explorer (The University of Manchester) (submittedVersion)
- Funders
- Wellcome Trust
- Royal Society
- Medical Research Council
- Biotechnology and Biological Sciences Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Health Sciences › Medicine › Oncology › Viral-associated cancers and disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- Epstein-Barr virus, Zta, BRLF1, epigenetic silencing, DNA methylation, CpG motifs, bZIP transcription factors, cysteine residues, serine residues, Burkitt lymphoma, viral latency, protein-DNA interactions
- MeSH terms
- Base Sequence, Cysteine, DNA-Binding Proteins, Herpesvirus 4, Human, Humans, Molecular Sequence Data, Serine, Viral Proteins, Trans-Activators, Gene Expression Regulation, Viral, Genome, Viral, DNA Methylation, Mutation, Missense, Response Elements, Gene Silencing, Epigenesis, Genetic, Cell Line, Tumor
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