ATP-independent Control of Vac8 Palmitoylation by a SNARE Subcomplex on Yeast Vacuoles
Abstract
Yeast vacuole fusion requires palmitoylated Vac8. We previously showed that Vac8 acylation occurs early in the fusion reaction, is blocked by antibodies against Sec18 (yeast N-ethylmaleimide-sensitive fusion protein (NSF)), and is mediated by the R-SNARE Ykt6. Here we analyzed the regulation of this reaction on purified vacuoles. We show that Vac8 acylation is restricted to a narrow time window, is independent of ATP hydrolysis by Sec18, and is stimulated by the ion chelator EDTA. Analysis of vacuole protein complexes indicated that Ykt6 is part of a complex distinct from the second R-SNARE, Nyv1. We speculate that during vacuole fusion, Nyv1 is the classical R-SNARE, whereas the Ykt6-containing complex has a novel function in Vac8 palmitoylation.
Additional Information
© 2005 the American Society for Biochemistry and Molecular Biology. Received for publication, September 14, 2004, and in revised form, January 18, 2005. Originally published In Press as doi:10.1074/jbc.M410582200 on February 10, 2005. Originally published In Press as doi:10.1074/jbc.M410582200 on February 8, 2005. We thank Alan Morgan for the Sec18 E350Q mutant, Walter Nickel and Rico Laage for plasmids and strains, and Nadine Decker and Gabriela Müller for expert technical assistance. This work was supported by Grant UN 111/2-3 from the Deutsche Forschungsgemeinschaft, by SFB638, the European Molecular Biology Organization Young Investigator Programme, and the Fonds der Chemischen Industrie (to C.U.), by predoctoral fellowships from the Boehringer Ingelheim Fonds (to L.E.P.D.) and the National Science Foundation Graduate Research Fellowship Program, and by a Chica & Heinz Schaller Stiftung/Deutscher Akademischer Austauschdienst University of Heidelberg Molecular and Cellular Biology Programme Fellowship (to T.J.L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.Attached Files
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Identifiers
- Eprint ID
- 5958
- Resolver ID
- CaltechAUTHORS:DIEjbc05
Funding
- Deutsche Forschungsgemeinschaft (DFG)
- UN 111/2-3
- Deutsche Forschungsgemeinschaft (DFG)
- SFB638
- European Molecular Biology Organization (EMBO)
- Fonds der Chemischen Industrie
- Boehringer Ingelheim Fonds
- NSF Graduate Research Fellowship
- Chica and Heinz Schaller Stiftung
- Deutscher Akademischer Austauschdienst (DAAD)
Dates
- Created
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2006-11-09Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field