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J. Biol. Chem., Vol. 277, Issue 16, 13831-13839, April 19, 2002
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From the Department of Biochemistry and Molecular Biology, State
University of New York, Upstate Medical University, Syracuse, New York
13210
Vacuolar proton-translocating ATPases are
composed of a peripheral complex, V1, attached to an
integral membrane complex, Vo. 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 V1 complexes released
from the membrane by glucose deprivation but binds constitutively to
cytosolic V1 sectors in a mutant lacking Vo
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·V1 interactions are specifically
disrupted in cells lacking V1 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·V1 binding; thus, Skp1p
recruitment to the RAVE complex does not appear to provide a signal for
V-ATPase assembly.
The RAVE Complex Is Essential for Stable Assembly of the Yeast
V-ATPase*
*
This work was supported by National Institutes of Health
Grant GM50322 (to P. M. K.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Biochemistry
and Molecular Biology, State University of New York, Upstate Medical
University, 750 East Adams St., Syracuse, NY 13210. Tel.: 315-464-8742;
Fax: 315-464-8736; E-mail: kanepm@upstate.edu.
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