CpG-Methylation Regulates a Class of Epstein-Barr Virus Promoters

Bergbauer, Martin; Kalla, Markus; Schmeinck, Anne; Göbel, Christine; Rothbauer, Ulrich; Eck, Sebastian; Benet‐Pagès, Anna; Strom, Tim M.; Hammerschmidt, Wolfgang

doi:10.1371/journal.ppat.1001114

Cited by 99 publications

(162 citation statements)

References 59 publications

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“…5A). The results indicated that viral transcription preferentially occurred from CpG-methylated virion template DNA as predicted (4). The ratio between DOXtreated meEBV-infected Daudi cells and wtEBV-infected cells without DOX corroborated this earlier finding at the level of individual genes, as can be seen with transcripts of four genes which were induced 400-to 500-fold (BRLF1, BMRF1), 350-fold (BcLF1), and more than 1,000-fold (BLLF1-gp220).…”

Section: Latently Infected Ebv-positive Akata Cells Support the Lytimentioning

confidence: 56%

“…In general, herpesviral virion DNA is free of 5-methylcytosines, which is surprising in the case of EBV because its lytic DNA replication relies on highly CpG-methylated viral DNA in latently infected cells (4,26). In proliferating somatic cells, methylated cytosine residues are faithfully preserved during DNA replication and copied onto newly synthesized DNA strands, but our findings Daudi cells stably transfected with the DOX-regulated conditional expression plasmid iBZLF1 were infected with wtEBV and meEBV virus stocks for 2 days and incubated for two additional days with or without DOX.…”

Section: Discussionmentioning

confidence: 99%

“…However, BZLF1 transactivates certain early genes of EBV only when their promoters contain CpG-methylated BZLF1 binding sites (4). A number of early viral genes regulated in this manner code for viral factors and enzymes indispensable for lytic DNA replication of EBV (16).…”

Section: Generation and Quantification Of Virions With Unmethylatedmentioning

confidence: 99%

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The Lytic Phase of Epstein-Barr Virus Requires a Viral Genome with 5-Methylcytosine Residues in CpG Sites

Kalla

Göbel

Hammerschmidt

2012

J Virol

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Epstein-Barr virus (EBV) is a human herpesvirus which has been studied intensively for its role in certain human tumors. It also serves as a model of herpesviral latency because it establishes an immediate, latent infection in human B cells. When EBV infects quiescent, primary B cells it induces their continuous proliferation to yield growth-transformed B-cell lines in vitro. The lytic or productive phase of EBV's life cycle is induced by the expression of the viral BZLF1 gene in latently infected cells. The BZLF1 protein is a transactivator, which selectively binds to two classes of distinct DNA sequence motifs. One class is similar to the motifs that are bound by members of the AP-1 transcription factor family to which BZLF1 belongs. The second class, which contains CpG motifs, is predominant in viral promoters of early lytic genes and is BZLF1's preferred or exclusive target sequence when methylated. The BZLF1 gene is transiently expressed in newly infected B cells but fails to induce EBV's lytic cycle, potentially because the virion DNA is unmethylated. Here we report that the lack of 5-methylcytosine residues in CpG sites of virion DNA prevents the expression of essential lytic genes indispensable for viral DNA amplification during productive infection. This finding indicates that BZLF1 transactivates these promoters in a methylation-dependent fashion and explains how progeny virus synthesis is abrogated in newly infected B cells. Our data also reveal that viral lytic DNA synthesis precludes CpG methylation of virion DNA during EBV's lytic, productive cycle, which can be overcome by the ectopic expression of a prokaryotic cytosine methyltransferase to yield CpG-methylated virion DNA. Upon infection of B cells, randomly CpG-methylated virion DNA induces high expression of essential lytic genes in contrast to virion DNA free of 5-methylcytosine residues. Our data suggest that unmethylated virion DNA is part of EBV's strategy to prevent the viral lytic phase in newly infected B cells, allowing it to establish its characteristic latent infection in them.

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Section: Latently Infected Ebv-positive Akata Cells Support the Lytimentioning

confidence: 56%

Section: Discussionmentioning

confidence: 99%

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The Lytic Phase of Epstein-Barr Virus Requires a Viral Genome with 5-Methylcytosine Residues in CpG Sites

Kalla

Göbel

Hammerschmidt

2012

J Virol

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“…We wanted to investigate if VLP-derived BZLF1 transcripts regulate downstream genes and chose the BMRF1 gene as a representative example. This gene is prominently up-regulated early after infection (8) presumably because its promoter contains several BZLF1-responsive elements (43). EBV-negative Daudi cells with a BMRF1 promoter-driven luciferase gene served as reporter for the early expression of BZLF1.…”

Section: Resultsmentioning

confidence: 99%

RNAs in Epstein–Barr virions control early steps of infection

Jochum

Ruiss

Moosmann

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Herpesviruses are dsDNA viruses, but their virions may additionally contain RNAs that can be transduced to recipient cells. The biological functions of herpes virion RNA species are unknown. Here we address this issue for EBV, a widespread human herpesvirus with oncogenic potential. We show that EBV-derived particles that include virions, virus-like particles, and subviral vesicles contain viral mRNAs, microRNAs, and other noncoding RNAs. Viral RNAs were transduced during infection and deployed immediate functions that enhanced EBV’s capacity to transform primary B cells. Among these transduced viral RNAs, BZLF1 transcripts transactivated viral promoters triggering the prelatent phase of EBV infection, noncoding EBV-encoded RNA transcripts induced cellular cytokine synthesis, and BNLF2a mRNA led to immune evasion that prevented T-cell responses to newly infected B cells. Hence, transduced viral RNAs govern critical processes immediately after infection of B cells with EBV and likely play important roles in herpesviral infection in general.

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“…Routes by which Zta activates gene expression have been documented for both viral and host promoters. Many promoters are targeted by the interaction of the sequence-specific DNA-binding domain of Zta with sequence-specific 7 nt DNA elements, termed ZREs (Adamson & Kenney, 1999;Bergbauer et al, 2010;Bhende et al, 2004Bhende et al, , 2005Broderick et al, 2009;Dickerson et al, 2009;Flower et al, 2011;Holley-Guthrie et al, 1990;Kalla et al, 2012Kalla et al, , 2010Karlsson et al, 2008;Kenney et al, 1989;Ramasubramanyan et al, 2012a, b;Sinclair, 2003;Sinclair et al, 1991;Woellmer et al, 2012). At least 32 distinct ZRE sequence variants are specifically recognized by Zta (Flower et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Repression of CIITA by the Epstein–Barr virus transcription factor Zta is independent of its dimerization and DNA binding

Balan

Osborn

Sinclair

2016

Journal of General Virology

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Repression of the cellular CIITA gene is part of the immune evasion strategy of the cherpes virus Epstein-Barr virus (EBV) during its lytic replication cycle in B-cells. In part, this is mediated through downregulation of MHC class II gene expression via the targeted repression of CIITA, the cellular master regulator of MHC class II gene expression. This repression is achieved through a reduction in CIITA promoter activity, initiated by the EBV transcription and replication factor, Zta (BZLF1, EB1, ZEBRA). Zta is the earliest gene expressed during the lytic replication cycle. Zta interacts with sequence-specific elements in promoters, enhancers and the replication origin (ZREs), and also modulates gene expression through interaction with cellular transcription factors and co-activators. Here, we explore the requirements for Zta-mediated repression of the CIITA promoter. We find that repression by Zta is specific for the CIITA promoter and can be achieved in the absence of other EBV genes. Surprisingly, we find that the dimerization region of Zta is not required to mediate repression. This contrasts with an obligate requirement of this region to correctly orientate the DNA contact regions of Zta to mediate activation of gene expression through ZREs. Additional support for the model that Zta represses the CIITA promoter without direct DNA binding comes from promoter mapping that shows that repression does not require the presence of a ZRE in the CIITA promoter.

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CpG-Methylation Regulates a Class of Epstein-Barr Virus Promoters

Cited by 99 publications

References 59 publications

The Lytic Phase of Epstein-Barr Virus Requires a Viral Genome with 5-Methylcytosine Residues in CpG Sites

The Lytic Phase of Epstein-Barr Virus Requires a Viral Genome with 5-Methylcytosine Residues in CpG Sites

RNAs in Epstein–Barr virions control early steps of infection

Repression of CIITA by the Epstein–Barr virus transcription factor Zta is independent of its dimerization and DNA binding

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