Glucosylceramide synthases, a gene family responsible for the biosynthesis of glucosphingolipids in animals, plants, and fungi.
Glucosylceramides are membrane lipids in most eukaryotic organisms and in a few bacteria. The physiological functions of these glycolipids have only been documented in mammalian cells, whereas very little information is available of their roles in plants, fungi, and bacteria. In an attempt to establish appropriate experimental systems to study glucosylceramide functions in these organisms, we performed a systematic functional analysis of a glycosyltransferase gene family with members of animal, plant, fungal, and bacterial origin. Deletion of such putative glycosyltransferase genes in Candida albicans and Pichia pastoris resulted in the complete loss of glucosylceramides. When the corresponding knock-out strains were used as host cells for homologous or heterologous expression of candidate glycosyltransferase genes, five novel glucosylceramide synthase (UDP-glucose:ceramide glucosyltransferase) genes were identified from the plant Gossypium arboreum (cotton), the nematode Caenorhabditis elegans, and the fungi Magnaporthe grisea, Candida albicans, and P. pastoris. The glycosyltransferase gene expressions led to the biosynthesis of different molecular species of glucosylceramides that contained either C18 or very long chain fatty acids. The latter are usually channeled exclusively into inositol-containing sphingolipids known from Saccharomyces cerevisiae and other yeasts. Implications for the biosynthesis, transport, and function of sphingolipids will be discussed.
Leipelt
M
M
Institut für Allgemeine Botanik, University of Hamburg, Ohnhorststr. 18, 22609 Hamburg, Germany.
Warnecke
D
D
Zähringer
U
U
Ott
C
C
Müller
F
F
Hube
B
B
Heinz
E
E
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D017173:Caenorhabditis elegans / Q000201:enzymology; D002176:Candida albicans / Q000201:enzymology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D005963:Glucosylceramides / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics*; D003368:Gossypium / Q000201:enzymology; D006801:Humans; D008055:Lipids / Q000737:chemistry; D020082:Magnaporthe / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008954:Models, Biological; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D010843:Pichia / Q000201:enzymology; D010940:Plant Proteins / Q000737:chemistry; D030821:Plants, Genetically Modified; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D013107:Sphingolipids / Q000737:chemistry*
D018076:DNA, Complementary; D005656:Fungal Proteins; D005963:Glucosylceramides; D008055:Lipids; D010940:Plant Proteins; D013107:Sphingolipids; D005964:Glucosyltransferases; C023246:ceramide glucosyltransferase