2. Academic Validation
  3. A heterotrimeric protein complex assembles the metazoan V-ATPase upon dissipation of proton gradients

A heterotrimeric protein complex assembles the metazoan V-ATPase upon dissipation of proton gradients

  • Nat Struct Mol Biol. 2025 Jul 11. doi: 10.1038/s41594-025-01610-9.
Christopher Nardone 1 2,  Julian Mintseris 3,  Dingwei He 3 4,  Justine C Rutter 5 6,  Benjamin L Ebert 5 6,  Steven P Gygi 3,  Tom Rapoport 7 8
Affiliations

Affiliations

  • 1 Department of Cell Biology, Harvard Medical School, Boston, MA, USA. christopher_nardone@hms.harvard.edu.
  • 2 Howard Hughes Medical Institute, Boston, MA, USA. christopher_nardone@hms.harvard.edu.
  • 3 Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
  • 4 Howard Hughes Medical Institute, Boston, MA, USA.
  • 5 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
  • 6 Cancer Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • 7 Department of Cell Biology, Harvard Medical School, Boston, MA, USA. tom_rapoport@hms.harvard.edu.
  • 8 Howard Hughes Medical Institute, Boston, MA, USA. tom_rapoport@hms.harvard.edu.
PMID: 40646309 DOI: 10.1038/s41594-025-01610-9
Abstract

Organelles such as lysosomes and synaptic vesicles are acidified by V-ATPases, which consist of a cytosolically oriented V1 complex that hydrolyzes ATP and a membrane-embedded VO complex that pumps protons. In yeast, V1-VO association is facilitated by the RAVE (regulator of H+-ATPase of the vacuolar and endosomal membrane) complex, but how higher eukaryotes assemble V-ATPases remains unclear. Here we identify a metazoan RAVE complex (mRAVE) whose structure and composition are notably divergent from the ancestral counterpart. mRAVE consists of DMXL1 or DMXL2, WDR7 and the central linker ROGDI. DMXL1 and DMXL2 interact with subunits A and D of the inactive, isolated V1. On dissipation of proton gradients, mRAVE binds to V1 and VO, forming a supercomplex on the membrane. mRAVE then catalyzes V1-VO assembly, enabling lysosomal acidification, neurotransmitter loading into vesicles and ATG16L1 recruitment for LC3/ATG8 conjugation onto single membranes. Our findings provide a molecular basis for neurological disorders caused by mRAVE mutations.

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