ATG14 is an essential regulator of autophagy that localizes the PI3-kinase complex PI3KC3-C1 to initiation sites and drives multiple stages of autophagosomal biogenesis. It promotes autophagosome formation by facilitating MAP1LC3/LC3 conjugation to phosphatidylethanolamine and directing BECN1 translocation from the trans-Golgi network to autophagosomes. ATG14 also binds the STX17-SNAP29 t-SNARE complex on autophagosomes to prime autophagosome-lysosome fusion. During starvation, ATG14 relocalizes to ER-mitochondria contact sites where autophagosome biogenesis occurs 1. ATG14 is critical for both canonical and non-canonical autophagy; deletion of ATG14 in human macrophages increases Mycobacterium tuberculosis replication by impairing phagosomal maturation 2. In atherosclerosis, ATG14 expression is reduced in macrophages; augmenting ATG14 restores autophagosome-lysosome fusion, promotes lipid degradation, and reduces atherosclerotic lesion formation 3. ATG14 elevation also drives chemoresistance in gastric cancer by increasing autophagy 4, and regulates hypoxia-induced autophagy in hepatocellular carcinoma 5. ATG14-mediated autophagy additionally contributes to protective mitochondrial quality control during cardiac ischemia 6 and oocyte aging 7. These findings implicate ATG14 as a therapeutic target for modulating autophagy in cardiovascular disease, cancer, and infection.