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Originally published In Press as doi:10.1074/jbc.M200682200 on February 13, 2002

J. Biol. Chem., Vol. 277, Issue 16, 13831-13839, April 19, 2002
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The RAVE Complex Is Essential for Stable Assembly of the Yeast V-ATPase*

Anne M. Smardon, Maureen Tarsio, and Patricia M. KaneDagger

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.


* 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.

Dagger 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.


Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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