Epstein-Barr virus (EBV) is a prevalent human herpesvirus affecting over 90% of the adult population worldwide.
To observe EBV-dependent epigenetic changes in epithelial cells, our lab infected immortalized normal oral keratinocytes (NOKs) with EBV for several passages and then allowed infected cells to naturally lose the virus (transient infection). To correlate DNA methylation to functional changes in gene expression, the transcriptional profiles following transient EBV infection were analyzed compared to uninfected parental controls. In conclusion, our observations suggest that EBV infection of epithelial cells results in epigenetic changes to the host genome that have long lasting phenotypic consequences even after loss of the virus, and define a potential mechanism for viral “hit-and-run” oncogenesis. Related PostsEpigenetic Regulation is Involved in Host Antiviral Defense MechanismsWanna Be Forever 21?
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Enter your email address to subscribe to EpiBeat and receive notifications of new posts by email. If EBV induced epigenetic changes, these epigenetic alterations would be retained in clones that lost EBV.
Hino R, Uozaki H, Murakami N, Ushiku T, Shinozaki A, Ishikawa S, Morikawa T, Nakaya T, Sakatani T, Takada K, & Fukayama M (2009).
Activation of DNA methyltransferase 1 by EBV LMP1 Involves c-Jun NH(2)-terminal kinase signaling.


The Epstein-Barr virus oncogene product, latent membrane protein 1, induces the downregulation of E-cadherin gene expression via activation of DNA methyltransferases.
Global and non-random CpG-island methylation in gastric carcinoma associated with Epstein-Barr virus.
Epigenetic markers for early detection of nasopharyngeal carcinoma in a high risk population. Genome-wide DNA methylation as an epigenetic consequence of Epstein-Barr virus infection of immortalized keratinocytes.
EBV infected NOKs showed a delay in differentiation in response to calcium and methylcellulose, two well established methods of inducing keratinocyte differentiation. Activation of DNA methyltransferase 1 by EBV latent membrane protein 2A leads to promoter hypermethylation of PTEN gene in gastric carcinoma. The viral life cycle is dependent on host epigenetic machinery to regulate viral gene expression programs that ensure viral persistence and spread. Importantly, this phenotype was maintained in EBV-negative transiently infected cells, suggestive of epigenetic alterations. Downregulated genes acquired DNA methylation typically near the transcription start site (TSS) while upregulated genes acquired DNA methylation much farther from the TSS. Given that EBV can modulate the DNMTs, we used reduced representation bisulfite sequencing to map the methylation status of CpG residues genome wide.


Several EBV proteins interact with host epigenetic machinery, such as the latent membrane proteins ability to induce the activity of the DNA methyltransferases (DMNT) [1-3]. Treatment with a DNMT inhibitor, decitabine, increased RNA levels in a set of downregulated genes similar to RNA levels in uninfected controls, suggesting that the increased DNA methylation was responsible for the altered regulation [6]. The consequences of which can best be seen in EBV-associated malignancies with the CpG island methylator phenotype (CIMP) [4, 5]. Further analysis revealed over 13,000 differentially methylated CpGs that were common in cells exposed to EBV compared to uninfected parental controls.
CIMP is where CpG islands upstream of tumor suppressors are hypermethylated and this phenotype has a strong correlation with EBV. Of particular interest was the increase in methylation at CpG islands associated with 27 genes in cells exposed to EBV compared to uninfected controls, a phenomenon reminiscent of the CIMP phenotype seen in vivo in EBV-associated carcinomas.
Whether EBV directs the DNA methylation changes, or if the epigenetic changes are due to the carcinogenic process are still being defined.



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