The impact of Influenza A Virus infection on m6A RNA in MDCK cells
m6A is the most common RNA modification in eukaryotic cells. Spatially ordered molecular interactions associated with m6A modification are crucial for the spatial organization and functional distribution in living cells. Our m6A proximity labeling technique utilizes a fusion protein containing the YTH domain to identify m6A sites in host cells before and after viral infection. The m6A-modified RNA is pulled down and subjected to high-throughput RNA sequencing. The results indicate that after infection with Influenza A Virus, the m6A modification levels of ribosome-associated mRNA in host cells are upregulated. By combining downstream molecular biology techniques, we have determined that the upregulation of m6A levels in ribosome-associated mRNA is a key factor in promoting viral infection of cells, which is of significant importance for the synthesis of Influenza A Virus proteins. Overall design: In MDCK cells, a fusion protein YTH-APEX2 containing the YTH domain was transfected for expression. The control group received no further treatment, while the experimental group was infected with the virus for 24 hours. Subsequently, m6A-modified RNA was pulled down using an antibody against the tag on the fusion protein for high-throughput sequencing analysis.