EDC4 C-terminal domain scaffolds P-body assembly and links P-body dynamics to p53 mediated tumor suppression

Processing bodies (P-bodies) are membrane-less organelles involved in mRNA metabolism, including decay, storage, and translational repression. Their formation is driven by liquid-liquid phase separation, primarily mediated by the scaffold protein EDC4. This study identifies the C-terminal domain of EDC4 as essential for P-body assembly. Introducing the microprotein NBDY selectively disrupted P-body formation by inhibiting EDC4 self-interaction, without affecting mRNA decay. Disruption of P-bodies led to upregulation of genes in the p53 signaling pathway, highlighting a link between P-body dynamics and tumor suppression. Clinical data suggest a possible connection between P-body dysregulation and cancer progression. Overall design: In this experiment, the microprotein NBDY (amino acids 22-41) was overexpressed in HEK293T cells to disrupt P-body formation. The aim was to investigate the functional consequences of P-body disruption. Following NBDY overexpression, RNA-seq and RT-qPCR were performed to analyze changes in gene expression, with a particular focus on pathways affected by P-body dynamics. The results were used to explore the broader role of P-bodies in cellular processes, including their involvement in tumor suppression mechanisms.