The V-ATPase/ATG16L1 axis is controlled by the V1H subunit

• DOI: 10.1016/j.molcel.2024.07.003
• Publication Date: 2024-07-31T14:39:02Z
• Authors: Lewis Timimi; Antoni G. Wrobel; George N. Chiduza; Sarah L. Maslen; Antonio Torres-Mé...; Beatriz Montaner; Colin Davis; Taylor Minckley; Katriona L. Hole; Andrea Serio; Michael J. Devine; J. Mark Skehel; John L. Rubinstein; Anne Schreiber; Rupert Beale
• Keywords: Neurons; Mice, Knockout; Vacuolar Proton-Translocating ATPases; FOS: Clinical medicine; Induced Pluripotent Stem Cells; Immunology; Autophagy-Related Proteins; Infectious Disease; Autophagy-Related Protein 8 Family; Cell Biology; Biochemistry & Proteomics; Mice, Inbred C57BL; Mice; HEK293 Cells; Influenza, Human; Autophagy; Humans; Animals; Carrier Proteins; Protein Binding; Signal Transduction

Defects in organellar acidification indicate compromised or infected compartments. Recruitment of the autophagy-related ATG16L1 complex to pathologically neutralized organelles targets ubiquitin-like ATG8 molecules to perturbed membranes. How this process is coupled to proton gradient disruption is unclear. Here, we reveal that the V1H subunit of the vacuolar ATPase (V-ATPase) proton pump binds directly to ATG16L1. The V1H/ATG16L1 interaction only occurs within fully assembled V-ATPases, allowing ATG16L1 recruitment to be coupled to increased V-ATPase assembly following organelle neutralization. Cells lacking V1H fail to target ATG8s during influenza infection or after activation of the immune receptor stimulator of interferon genes (STING). We identify a loop within V1H that mediates ATG16L1 binding. A neuronal V1H isoform lacks this loop and is associated with attenuated ATG8 targeting in response to ionophores in primary murine and human iPSC-derived neurons. Thus, V1H controls ATG16L1 recruitment following proton gradient dissipation, suggesting that the V-ATPase acts as a cell-intrinsic damage sensor.