Arginine methylation controls cell proliferation by integrating E2F activity with the splicing machinery (RIP-seq data set)

Residue-specific arginine methylation (meR) on the E2F1 subunit plays an essential role in determining whether cell cycle progression or apoptosis ensues, although the molecular pathways underpinning the effects of methylation remain obscure. We report here that the methylation events on E2F1 have a direct impact on the mechanisms which underpin gene expression control. Specifically, the PRMT5-mediated symR mark not only influences the transcription repertoire of genes targeted by E2F1, but also endows E2F1 with the ability to regulate RNA splicing. This occurs through tudor domain protein p100/TSN which reads the symR mark on E2F1, and thereby enables a distinct set of RNA to be alternatively spliced. The p100/TSN-E2F1 complex binds alternatively spliced RNAs, with many RNAs derived from E2F target genes connected with proliferation and cancer. Overall design: U2OS cells were transfected with siRNA targeting p100/TSN or non-targeting siRNA for 72h. Cells transfected with siE2F1 were used as a reference sample for specificity. An E2F-1 antibody was used in a RNA immunoprecipitation (RIP) reaction and RNA was extracted and DNAse treated from duplicate biological replicates. Samples underwent ribodepletion prior to sequencing on Illumina NextSeq.