The RAVE complex is essential for stable assembly of the yeast V-ATPase

J Biol Chem. 2002 Apr 19;277(16):13831-9. doi: 10.1074/jbc.M200682200. Epub 2002 Feb 13.

Abstract

Vacuolar proton-translocating ATPases are composed of a peripheral complex, V(1), attached to an integral membrane complex, V(o). Association of the two complexes is essential for ATP-driven proton transport and is regulated post-translationally in response to glucose concentration. A new complex, RAVE, was recently isolated and implicated in glucose-dependent reassembly of V-ATPase complexes that had disassembled in response to glucose deprivation (Seol, J. H., Shevchenko, A., and Deshaies, R. J. (2001) Nat. Cell Biol. 3, 384-391). Here, we provide evidence supporting a role for RAVE in reassembly of the V-ATPase but also demonstrate an essential role in V-ATPase assembly under other conditions. The RAVE complex associates reversibly with V(1) complexes released from the membrane by glucose deprivation but binds constitutively to cytosolic V(1) sectors in a mutant lacking V(o) sectors. V-ATPase complexes from cells lacking RAVE subunits show serious structural and functional defects even in glucose-grown cells or in combination with a mutation that blocks disassembly of the V-ATPase. RAVE small middle dotV(1) interactions are specifically disrupted in cells lacking V(1) subunits E or G, suggesting a direct involvement for these subunits in interaction of the two complexes. Skp1p, a RAVE subunit involved in many different signal transduction pathways, binds stably to other RAVE subunits under conditions that alter RAVE small middle dotV(1) binding; thus, Skp1p recruitment to the RAVE complex does not appear to provide a signal for V-ATPase assembly.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Cycle Proteins / metabolism
  • Cell Membrane / metabolism
  • Fungal Proteins / metabolism
  • Genotype
  • Glucose / metabolism
  • Models, Biological
  • Mutation
  • Precipitin Tests
  • Protein Binding
  • S-Phase Kinase-Associated Proteins
  • Signal Transduction
  • Vacuolar Proton-Translocating ATPases / chemistry*
  • Vacuolar Proton-Translocating ATPases / metabolism*

Substances

  • Cell Cycle Proteins
  • Fungal Proteins
  • S-Phase Kinase-Associated Proteins
  • Vacuolar Proton-Translocating ATPases
  • Glucose