METTL3 alters capping enzyme expression and its activity on ribosomal proteins in NSCLC cell model
The 5' cap, catalyzed by capping enzyme, RNGTT, is a vital mRNA modification for the functionality of mRNAs and dysregulated in cancer cells. The m6A writer METTL3 is a regulator of mRNA stability and protein translation, and its expression is altered in various cancers, including lung cancer cells. We identified a METTL3 motif in the mRNA of RNGTT and investigated whether METTL3 could regulate the expression of RNGTT. Knock down of METTL3 resulted in destabilizing RNGTT mRNA, reduced protein expression and activity. Sequencing of capped mRNAs and m7G immunoprecipitation identified an underrepresentation of ribosomal protein mRNA. Overall design: To investigate the regulatory relationship between METTL3 and RNGTT, we generated a stable cell line of A549 expressing either an sh- control or shMETTL3 We then performed a cDNA enrichment of capped mRNAs in biological triplicate of A549sh- and A549 shMETTL3. The then performed gene expression profile analysis using data obtained from DNAseq with the resulting cDNA.