Autophagy is dynamically regulated by nutrient status and intracellular signaling. Rubicon is a well-known negative regulator of autophagy, but the mechanism underlying its inhibition has remained unclear. Here we identify a novel Rubicon interactor that colocalizes with lysosomes and positively influences autophagic flux. Importantly, Rubicon depletion triggers lysosomal Ca²⁺ transients, which are required for autophagic activation through the TRPML1 channel. Disrupting either TRPML1 function or Ca²⁺ availability abolishes this response. These findings indicate that Rubicon and its novel interactor converge on a TRPML1-dependent Ca²⁺ signaling step shared with starvation-induced autophagy.