Conclusions
These results reveal the critical role of m6 A modification in the process of de novo EBV infection.
Methods
Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and MeRIP-RT-qPCR were used to determine the m6 A-modified transcripts during de novo EBV infection. RIP assay was used to confirm the binding of EBNA2 and m6 A readers. Quantitative reverse-transcription polymerase chain reaction (RT-qPCR) and Western blot analysis were performed to test the effect of m6 A on the host and viral gene expression.
Results
Here, we provided mechanistic insights by examining the viral and cellular m6 A epitranscriptomes during de novo EBV infection, which is in the pre-latent phase. EBV EBNA2 and BHRF1 were highly m6 A-modified upon EBV infection. Knockdown of METTL3 (a "writer") decreased EBNA2 expression levels. The emergent m6 A modifications induced by EBV infection preferentially distributed in 3' untranslated regions of cellular transcripts, while the lost m6 A modifications induced by EBV infection preferentially distributed in coding sequence regions of mRNAs. EBV infection could influence the host cellular m6 A epitranscriptome. Conclusions: These results reveal the critical role of m6 A modification in the process of de novo EBV infection.