The Ral small GTPase is an essential regulator of Exocyst complex function in secretion

ABSTRACT Ral GTPases have long been proposed as regulators of the metazoan Exocyst, a conserved secretory vesicle-tethering complex, but direct evidence for this role has been scarce. In contrast, the well-studied yeast Exocyst relies on multiple Rab GTPases to regulate function, but yeast do not encode Ral. Using Caenorhabditis elegans we demonstrate that endogenous RAL-1 directly engages the Exocyst through conserved binding sites in its subunits. Loss of RAL-1 disrupts dendritic arborization of PVD sensory neurons, impairs vesicle trafficking, and causes broad developmental defects, acting both cell-autonomously in neurons and non-autonomously through supporting epithelial cells. Structure-guided genome editing of RAL-1-Exocyst interfaces produced synthetic phenotypes, underscoring the physiological importance of these contacts. Taken together, our findings establish RAL-1 as a bona fide regulator of the metazoan Exocyst in vivo and suggest that Ral-Exocyst interactions operate in parallel with other secretory pathways. More broadly, this work positions C. elegans as a powerful system to dissect Ral- Exocyst mechanisms across molecular, cellular, and developmental scales.