ATP-independent control of Vac8 palmitoylation by a SNARE subcomplex on yeast vacuoles.
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.
Dietrich
Lars E P
LE
Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany.
LaGrassa
Tracy J
TJ
Rohde
Jan
J
Cristodero
Marina
M
Meiringer
Christoph T A
CT
Ungermann
Christian
C
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D005982:Glutathione Transferase / Q000378:metabolism; D006868:Hydrolysis; D047468:Immunoprecipitation; D008074:Lipoproteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D008858:Microscopy, Phase-Contrast; D008954:Models, Biological; D019308:Palmitic Acid / Q000378:metabolism; D011485:Protein Binding; D050683:R-SNARE Proteins; D011994:Recombinant Proteins / Q000737:chemistry; D050600:SNARE Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D013997:Time Factors; D014617:Vacuoles / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism*
D008074:Lipoproteins; D008565:Membrane Proteins; C500806:Nyv1 protein, S cerevisiae; D050683:R-SNARE Proteins; D011994:Recombinant Proteins; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C111373:VAC8 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C107127:YKT6 protein, S cerevisiae; D019308:Palmitic Acid; D000255:Adenosine Triphosphate; D004492:Edetic Acid; D005982:Glutathione Transferase; D000251:Adenosine Triphosphatases; C064404:SEC18 protein, S cerevisiae