The autophagy-lysosome pathway, governed by transcription factor EB (TFEB), is critical in cellular degradation and signal transduction. Dysregulation of this pathway or TFEB contributes to aging, tumorigenesis, and metabolic disorders. In this study, we observed that TFEB exhibits an initial activation (via dephosphorylation) followed by protein downregulation under prolonged stress conditions. This rapid termination of TFEB activation likely functions as a protective mechanism to prevent tumor cell death. Mechanistic studies revealed that vacuolar protein sorting-associated protein 11 (VPS11)—a downstream target gene of TFEB and an E3 ubiquitin ligase, is a negative regulator of TFEB. VPS11 interacts with TFEB through its C-terminal CHCR2 and RING domains, promoting TFEB degradation via K48-linked ubiquitination at lysine 264 (K264). Proteomic and functional analyses demonstrated that VPS11 knockdown activates autophagy while impairing autolysosome maturation—an effect significantly amplified by TFEB overexpression. Meanwhile, VPS11 was also found to transcriptionally regulate the expression of major histocompatibility complex class I (MHC-I) in a TFEB-dependent manner. Therefore, targeting VPS11 improves tumor antigen presentation and enhances the therapeutic efficacy of PD-1 blockade. Collectively, VPS11 is identified to be a novel negative regulator of TFEB, which orchestrates autolysosome pathway homeostasis and antigen presentation, and thereby modulates anti-tumor efficacy.