Ceramide plays a crucial role as a basic building block of sphingolipids, but also as a signalling molecule mediating cell-fate decisions. Three genes, LAG1, LAC1 and LIP1, have been shown to be required for ceramide synthase activity in Saccharomyces cerevisiae [Guillas, Kirchman, Chuard, Pfefferli, Jiang, Jazwinski and Conzelman (2001) EMBO J. 20, 2655–2665; Schorling, Vallee, Barz, Reizman and Oesterhelt (2001) Mol. Biol. Cell 12, 3417–3427; Vallee and Riezman (2005) EMBO J. 24, 730–741]. In the present study, the topology of the Lag1p and Lac1p subunits was investigated. The N- and C-termini of the proteins are in the cytoplasm and eight putative membrane-spanning domains were identified in Lag1p and Lac1p by insertion of glycosylation and factor Xa cleavage sites at various positions. The conserved Lag motif, potentially containing the active site, is most likely embedded in the membrane. We also present evidence that histidine and aspartic acid residues in the Lag motif are essential for the function of Lag1p in vivo.
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Research Article|
August 29 2006
Transmembrane topology of ceramide synthase in yeast
Natsuko Kageyama-Yahara;
Natsuko Kageyama-Yahara
1Department of Biochemistry, University of Geneva, Sciences II, 30, quai E. Ansermet, CH-1211 Geneva 4, Switzerland
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Howard Riezman
Howard Riezman
1
1Department of Biochemistry, University of Geneva, Sciences II, 30, quai E. Ansermet, CH-1211 Geneva 4, Switzerland
1To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
May 11 2006
Revision Received:
June 01 2006
Accepted:
June 07 2006
Accepted Manuscript online:
June 07 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 398 (3): 585–593.
Article history
Received:
May 11 2006
Revision Received:
June 01 2006
Accepted:
June 07 2006
Accepted Manuscript online:
June 07 2006
Citation
Natsuko Kageyama-Yahara, Howard Riezman; Transmembrane topology of ceramide synthase in yeast. Biochem J 15 September 2006; 398 (3): 585–593. doi: https://doi.org/10.1042/BJ20060697
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