RNA m⁶A methylation regulates virus–host interaction and EBNA2 expression during Epstein–Barr virus infection

Abstract

Introduction N6‐methyladenosine (m6A) is the most prevalent modification that occurs in messenger RNA (mRNA), affecting mRNA splicing, translation, and stability. This modification is reversible, and its related biological functions are mediated by “writers,” “erasers,” and “readers.” The field of viral epitranscriptomics and the role of m6A modification in virus–host interaction have attracted much attention recently. When Epstein–Barr virus (EBV) infects a human B lymphocyte, it goes through three phases: the pre‐latent phase, latent phase, and lytic phase. Little is known about the viral and cellular m6A epitranscriptomes in EBV infection, especially in the pre‐latent phase during de novo infection. Methods Methylated RNA immunoprecipitation sequencing (MeRIP‐seq) and MeRIP‐RT‐qPCR were used to determine the m6A‐modified transcripts during de novo EBV infection. RIP assay was used to confirm the binding of EBNA2 and m6A readers. Quantitative reverse‐transcription polymerase chain reaction (RT‐qPCR) and Western blot analysis were performed to test the effect of m6A on the host and viral gene expression. Results Here, we provided mechanistic insights by examining the viral and cellular m6A epitranscriptomes during de novo EBV infection, which is in the pre‐latent phase. EBV EBNA2 and BHRF1 were highly m6A‐modified upon EBV infection. Knockdown of METTL3 (a “writer”) decreased EBNA2 expression levels. The emergent m6A modifications induced by EBV infection preferentially distributed in 3ʹ untranslated regions of cellular transcripts, while the lost m6A modifications induced by EBV infection preferentially distributed in coding sequence regions of mRNAs. EBV infection could influence the host cellular m6A epitranscriptome. Conclusions These results reveal the critical role of m6A modification in the process of de novo EBV infection.

N6-methyladenosine regulates ATM expression and downstream signaling

“…From our previous results 26 , we found that the m6A abundance was increased in ATM mRNA transcripts upon EBV infection (Figure 1 A). ATM is a key kinase in DNA damage response, which is responsible for detecting, transmitting and repairing DNA damage response 21 .…”

“…Previous studies 23 - 25 including ours 26 have identified the effect of Epstein-Barr Virus (EBV) infection on the level of transcriptomic m6A modifications in host cells. Upon EBV infection, we identified increased levels of m6A modification of ATM mRNA.…”

Role of Virus-Induced Host Cell Epigenetic Changes in Cancer

“…Two recent studies reported that the EBV infection induces changes in N 6 -methyladenosine RNA methylation of viral and host cell mRNA. These epitransciptomic changes promoted EBV infection in vitro [ 391 , 392 ]. Once more, viruses take advantages of cellular processes to favor their life cycle.…”

The Biological Function, Mechanism, and Clinical Significance of m6A RNA Modifications in Head and Neck Carcinoma: A Systematic Review

“…Although the m6A modification was identified during the latent stage of EBV infection, the modification of EBV transcripts by the deposition of m6A was reduced during lytic infection. Increased EBNA3C levels have been shown to increase the expression and stability of METTL14, which can induce the proliferation and colony formation of EBV-positive cells ( Zheng et al, 2021 ) and stabilize METTL3 via its interaction with RNAs ( Klungland and Dahl, 2014 ; He and He, 2021 ). METTL3 is highly expressed in nasopharyngeal carcinoma tissues and affects the overall survival of patients with nasopharyngeal carcinoma.…”