Deciphering functional roles and interplay between Beclin1 and Beclin2 in autophagosome formation and mitophagy

The degradation of macromolecules and organelles by the process of autophagy is critical for cellular homeo-stasis and is often compromised during aging and disease. Beclin1 and Beclin2 are implicated in autophagy induction, and these homologs share a high degree of amino acid sequence similarity but have divergent N-terminal regions. Here, we investigated the functions of the Beclin homologs in regulating autophagy and mi-tophagy, a specialized form of autophagy that targets mitochondria. Both Beclin homologs contributed to au-tophagosome formation, but a mechanism of autophagosome formation independent of either Beclin homolog occurred in response to starvation or mitochondrial damage. Mitophagy was compromised only in Beclin1-de-ficient HeLa cells and mouse embryonic fibroblasts because of defective autophagosomal engulfment of mito-chondria, and the function of Beclin1 in mitophagy required the phosphorylation of the conserved Ser15 residue by the kinase Ulk1. Mitochondria-ER-associated membranes (MAMs) are important sites of autophagosome for-mation during mitophagy, and Beclin1, but not Beclin2 or a Beclin1 mutant that could not be phosphorylated at Ser15, localized to MAMs during mitophagy. Our findings establish a regulatory role for Beclin1 in selective mi-tophagy by initiating autophagosome formation adjacent to mitochondria, a function facilitated by Ulk1-me-diated phosphorylation of Ser15 in its distinct N-terminal region.