Acyl-CoA-binding protein, Acb1p, is required for normal vacuole function and ceramide synthesis in Saccharomyces cerevisiae

In the present study, we show that depletion of acyl-CoA-binding protein, Acb1p, in yeast affects ceramide levels, protein trafficking, vacuole fusion and structure. Vacuoles in Acb1p-depleted cells are multi-lobed, contain significantly less of the SNAREs (soluble N -ethylmaleimide-sensitive fusion...

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Published in:Biochemical journal 2004-06, Vol.380 (Pt 3), p.907-918
Main Authors: Faergeman, Nils J, Feddersen, Søren, Christiansen, Janne K, Larsen, Morten K, Schneiter, Roger, Ungermann, Christian, Mutenda, Kudzai, Roepstorff, Peter, Knudsen, Jens
Format: Article
Language:English
Subjects:
Carrier Proteins - physiology
Ceramides - biosynthesis
Ceramides - deficiency
Ceramides - metabolism
Diazepam Binding Inhibitor - metabolism
Membrane Fusion - physiology
Mutation - physiology
Protein Transport - physiology
Saccharomyces cerevisiae
Saccharomyces cerevisiae - chemistry
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins - metabolism
Saccharomyces cerevisiae Proteins - physiology
Spectrometry, Mass, Electrospray Ionization - methods
Vacuoles - chemistry
Vacuoles - metabolism
Vacuoles - physiology
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Summary:In the present study, we show that depletion of acyl-CoA-binding protein, Acb1p, in yeast affects ceramide levels, protein trafficking, vacuole fusion and structure. Vacuoles in Acb1p-depleted cells are multi-lobed, contain significantly less of the SNAREs (soluble N -ethylmaleimide-sensitive fusion protein attachment protein receptors) Nyv1p, Vam3p and Vti1p, and are unable to fuse in vitro. Mass spectrometric analysis revealed a dramatic reduction in the content of ceramides in whole-cell lipids and in vacuoles isolated from Acb1p-depleted cells. Maturation of yeast aminopeptidase I and carboxypeptidase Y is slightly delayed in Acb1p-depleted cells, whereas the maturation of alkaline phosphatase and Gas1p is unaffected. The fact that Gas1p maturation is unaffected by Acb1p depletion, despite the lowered ceramide content in these cells, indicates that ceramide synthesis in yeast could be compartmentalized. We suggest that the reduced ceramide synthesis in Acb1p-depleted cells leads to severely altered vacuole morphology, perturbed vacuole assembly and strong inhibition of homotypic vacuole fusion.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj20031949