RBM5 recruiting MGC32805 in a sandwich mode and inducing ΔFAS neoantigen and triggering FAS properties switch: implication in colorectal cancer.

Abstract

Pre-mRNA alternative splicing (AS) is a crucial process, which plays a significant role in inducing tumor subtype-specific alterations and the hallmark of epigenetic heterogeneity in tumorigenesis. However, the regulatory mechanisms of pre-mRNA AS remain obscure. This study demonstrates that splicing factor RBM5 recruits long non-coding RNA MGC32805, and they act in concert as oncogenes in colorectal cancer (CRC) cells by preventing apoptosis, as well as promoting migration and resistance to 5-Fluorouracil (5-FU). Specifically, they promote the exclusion of exon 6 in the FAS pre-mRNA, leading to decreased expression of mFAS (an apoptotic isoform) and increased expression of ΔFAS (an anti-apoptotic isoform) in both CRC cells and a mouse xenograft model. RBM5, which contains Leu650 and Arg681 residues in the ZnF-C2H2 domain, recognizes the "GUACG" (-1299 to -1303) motif in MGC32805. Furthermore, MGC32805 blocks the binding site (Lys645) of the E3 ubiquitin ligase PRPF19, which targets RBM5 for degradation, thus increasing the stability of RBM5. The His665 and Leu668 residues of RBM5 specifically bind to the FAS exon 6 adjacent element (GAACAAA), which drives FAS-AS events and increases the expression ratio of the ΔFAS/mFAS isoforms. These findings introduce a novel research strategy to investigate the epigenetic heterogeneity and plasticity of tumorigenesis. They also shed light on the mechanism of MGC32805-mediated transformation of the FAS tumor neoantigen function from a tumor suppressor to an oncogene at the AS level through its interactions with RBM5.