The E3 ligase MEX3B forms a tripartite complex with <i>Rest</i> and <i>Hotair</i> to determine the proliferative capacity of neural progenitor cells

E3 ubiquitin ligases regulate the cellular proteome <i>via</i> proteasome-dependent protein degradation; however, there exist limited studies outlining their non-canonical functions. RNA-binding ubiquitin ligases (RBULs) represent a subset of E3 ligases that harbour RNA-binding domains, making them uniquely positioned to function as both RNA-binding proteins and E3 ligases. Our initial microarray screen for E3 ligases from mouse cortical neural progenitor cells identified MEX3B, a known RNA-binding ubiquitin ligase, to be differentially expressed. Here, we characterize the non-canonical role of MEX3B in the context of neural proliferation. We find that MEX3B is significantly reduced following the differentiation of neural progenitor cells (NPCs). The knockdown of MEX3B blocks the proliferative state of NPCs and leads to the enhancement of neurite length and dendrite branching. We observed that MEX3B regulates the stability of <i>Rest</i> mRNA in proliferative NPCs. Mechanistically, MEX3B interacts with <i>Rest</i> mRNA and the lncRNA <i>Hotair</i> to form a tripartite complex in the presence of basic fibroblast growth factor (bFGF). Loss of <i>Hotair</i> disrupts this complex; conversely, MEX3B RNAi significantly reduces <i>Hotair</i> abundance. <i>Rest</i> mRNA levels remain unaffected by <i>Hotair</i> knockdown, suggesting that the latter acts as a scaffold to facilitate bFGF-dependent MEX3B<i>-Rest</i> interaction in the MEX3B<i>-Rest-Hotair</i> tripartite axis. Our study demonstrates an RNA-driven post-transcriptional mechanism underlying NPC proliferation.