Chemical inhibition of Exon Junction Complex assembly impairs mRNA localization and neural stem cells ciliogenesis

Abstract

The Exon Junction Complex (EJC) is formed by the essential eIF4A3, MAGOH and Y14 core proteins. It is universally deposited during splicing at exon-exon junctions. The EJC is known to impact almost every post-transcriptional regulatory step throughout the life of mRNAs including their modifications, splicing, decay and trafficking. Its dysregulation leads to neurodevelopmental pathologies. Here we show that EJC-i, a compound known to block the ATPase activity of eIF4A3, inhibits de novo EJC assembly. EJC-i and targeted knockdown of either eIF4A3 or Y14 core EJC subunits lead to very similar phenotypes by impacting the destiny of mRNAs due to alterations in alternative splicing, nonsense-mediated mRNA decay, genome-wide m6A methylation and proper intracellular addressing of specific transcripts, in particular to the centrosome. Both EJC impairment methods disrupt the centrosome function that might be responsible for mitotic arrest at prometaphase. As a small molecule that readily diffuses into cells, EJC-i is a particularly attractive easy to use and versatile tool to investigate EJC functions in live cells or whole organism that are not prone to genetic manipulation. Indeed, this property was used to disrupt ciliogenesis in primary neural stem cells.