ATP-independent control of Vac8 palmitoylation by a SNARE subcomplex on yeast vacuoles |
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Authors: | Dietrich Lars E P LaGrassa Tracy J Rohde Jan Cristodero Marina Meiringer Christoph T A Ungermann Christian |
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Institution: | Biochemie-Zentrum der Universit?t Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany. |
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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. |
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