CG14435 (detour), the Drosophila ZNRF1 and 2 homologue, regulates autophagosome biogenesis critical for healthy aging

Autophagy is essential for animal development and homeostasis, and its deregulation is associated with numerous human pathologies. Here we identify a novel regulator of autophagy, the RING domain containing ubiquitin ligase CG14435 (detour). Depletion of CG14435 resulted in increased early-stage autophagic puncta, premature tissue contraction, and an accumulation of autophagy related proteins in young adults. Overexpression of CG14435 or the mammalian homologues, Zinc and RING finger (ZNRF) 1 and ZNRF2, increased late-stage autophagy puncta size. The ablation of ZNRF1 or ZNRF2 in mammalian cells increased basal autophagy. We identified CG14435 interacting proteins including HOPS complex subunits, deep orange (Vps18), Vps16A, and light (Vps41). Furthermore, in the presence of CG14435, dor ubiquitination was increased. The CG14435 mutant adults displayed premature aging, impaired motor function (neurodegeneration), and activation of innate immunity. Our findings reveal a novel role for CG14435 in autophagy, likely through regulation of HOPS complex, required for healthy aging.