ULK1 induces autophagy by phosphorylating Beclin-1 and activating Vps34 lipid kinase. This study reveals that ULK1 promotes autophagy by phosphorylating Beclin-1 at serine 14, thereby enhancing the activity of the ATG14L-containing Vps34 complex. This phosphorylation is essential for autophagy induction in mammals and is conserved in C. elegans. The study shows that ULK1 activates the autophagy-specific Vps34 complex, which is necessary for autophagy initiation. The results indicate that ULK1 is required for the activation of the pro-autophagic Vps34 complex but not for the repression of non-autophagic Vps34 complexes. Additionally, Beclin-1 phosphorylation by ULK1 is required for the activation of the ATG14L-bound Vps34 complex. The study also demonstrates that Beclin-1 phosphorylation is required for autophagy induction in response to amino acid starvation. The findings highlight the critical role of ULK1 in autophagy regulation and the molecular mechanism linking ULK1 to autophagy initiation. The study further shows that the phosphorylation site on Beclin-1 is conserved in C. elegans and is required for proper autophagy. These results provide a comprehensive understanding of the molecular mechanisms underlying autophagy induction by ULK1.ULK1 induces autophagy by phosphorylating Beclin-1 and activating Vps34 lipid kinase. This study reveals that ULK1 promotes autophagy by phosphorylating Beclin-1 at serine 14, thereby enhancing the activity of the ATG14L-containing Vps34 complex. This phosphorylation is essential for autophagy induction in mammals and is conserved in C. elegans. The study shows that ULK1 activates the autophagy-specific Vps34 complex, which is necessary for autophagy initiation. The results indicate that ULK1 is required for the activation of the pro-autophagic Vps34 complex but not for the repression of non-autophagic Vps34 complexes. Additionally, Beclin-1 phosphorylation by ULK1 is required for the activation of the ATG14L-bound Vps34 complex. The study also demonstrates that Beclin-1 phosphorylation is required for autophagy induction in response to amino acid starvation. The findings highlight the critical role of ULK1 in autophagy regulation and the molecular mechanism linking ULK1 to autophagy initiation. The study further shows that the phosphorylation site on Beclin-1 is conserved in C. elegans and is required for proper autophagy. These results provide a comprehensive understanding of the molecular mechanisms underlying autophagy induction by ULK1.