Fine-tuning m6A and METTL3 levels have profound impact on cellular proliferation and protein synthesis

Using Oxford Nanopore Direct RNA Sequencing, we performed transcriptome-wide profiling of m6A modifications in wild-type and METTL3 heterozygous knockout cells. Computational approaches were applied to identify differential m6A sites and estimate modification stoichiometry across transcripts. These data were integrated with functional analyses including proliferation assays, quantitative proteomics, phosphoproteomics, and global protein synthesis measurements to assess the biological consequences of altered m6A levels.

The study aims to determine how partial reductions in METTL3 activity reshape the m6A landscape and influence gene expression programs, particularly those related to translational control, growth signalling, and cancer cell proliferation. Our findings demonstrate that modest decreases in global m6A preferentially affect transcripts involved in translational regulation and are associated with increased translational output and proliferative capacity, whereas stronger pharmacological inhibition of METTL3 suppresses translation and cell growth.

Overall, this work provides insight into the dose-dependent effects of METTL3 and m6A RNA methylation on gene regulation, highlighting how partial disruption of the m6A machinery can reprogram translational pathways and contribute to tumour progression.