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Structure of yeast RAVE bound to a partial V1 complex.

Hanlin Wang1,2, Maureen Tarsio3, Patricia M Kane3

  • 1Molecular Medicine Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.

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The regulator of the ATPase of vacuoles and endosomes (RAVE) complex in yeast binds to the V1 subcomplex of vacuolar-type ATPases (V-ATPases). This interaction primes V1 for reassembly with the VO complex, regulating proton transport.

Keywords:
Cryo-EMRAVEV-ATPaseassemblyregulation

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Structural Biology

Background:

  • Vacuolar-type ATPases (V-ATPases) are crucial proton pumps for intracellular acidification in eukaryotes.
  • V-ATPases comprise soluble V1 and membrane-bound VO subcomplexes that reversibly dissociate to regulate activity.
  • The RAVE complex is essential for V1-VO reassembly, facilitating V-ATPase function.

Purpose of the Study:

  • To elucidate the structural basis of RAVE-mediated V1-VO reassembly.
  • To determine the interaction interface between yeast RAVE and the V1 subcomplex.
  • To understand how RAVE binding influences V1 conformation for VO association.

Main Methods:

  • Electron cryomicroscopy (cryo-EM) was employed to visualize the yeast RAVE-V1 complex.
  • Structural analysis focused on the L-shaped RAVE complex and its interaction with V1 subunits.
  • Comparative analysis with ATP synthase structures identified unique binding regions.

Main Results:

  • The cryo-EM structure revealed an L-shaped RAVE complex with specific subunit orientations (Rav2p towards membrane, Skp1p distant).
  • Only Rav1p directly interacts with V1, binding to a unique region on subunit A not present in ATP synthases.
  • The V1 complex, though partially dissociated (missing subunits), adopts a conformation conducive to VO binding.

Conclusions:

  • RAVE acts as a scaffold, positioning V1 for efficient reassembly with VO.
  • The interaction interface and conformational changes induced by RAVE are key to regulating V-ATPase proton pumping activity.
  • This structural insight provides a foundation for understanding V-ATPase regulation and potential therapeutic targeting.