Glycobiology, Vol 8, 761-770, Copyright © 1998 by Society for Glycobiology
E Canivenc-Gansel, I Imhof, F Reggiori, P Burda, A Conzelmann and A Benachour
Cells synthesize the GPI anchor carbohydrate core by successively adding
N-acetylglucosamine, three mannoses, and phosphoethanolamine (EtN-P) onto
phosphatidylinositol, thus forming the complete GPI precursor lipid which
is then added to proteins. Previously, we isolated a GPI deficient yeast
mutant accumulating a GPI intermediate containing only two mannoses,
suggesting that it has difficulty in adding the third, alpha1,2-linked Man
of GPI anchors. The mutant thus displays a similar phenotype as the
mammalian mutant cell line S1A-b having a mutation in the PIG-B gene. The
yeast mutant, herein named gpi10-1 , contains a mutation in YGL142C, a
yeast homolog of the human PIG-B. YGL142C predicts a highly hydrophobic
integral membrane protein which by sequence is related to ALG9, a yeast
gene required for adding Man in alpha1,2 linkage to N-glycans. Whereas
gpi10-1 cells grow at a normal rate and make normal amounts of GPI
proteins, the microsomes of gpi10-1 are completely unable to add the third
Man in an in vitro assay. Further analysis of the GPI intermediate
accumulating in gpi10 shows it to have the structure
Manalpha1-6(EtN-P-)Manalpha1-4GlcNalpha1- 6(acyl) Inositol-P-lipid. The
presence of EtN-P on the alpha1,4-linked Man of GPI anchors is typical of
mammalian and a few other organisms but had not been observed in yeast GPI
proteins. This additional EtN-P is not only found in the abnormal GPI
intermediate of gpi10-1 but is equally present on the complete GPI
precursor lipid of wild type cells. Thus, GPI biosynthesis in yeast and
mammals proceeds similarly and differs from the pathway described for
Trypanosoma brucei in several aspects.
ORIGINAL ARTICLES
GPI anchor biosynthesis in yeast: phosphoethanolamine is attached to the alpha1,4-linked mannose of the complete precursor glycophospholipid
Institute of Biochemistry, University of Fribourg, Perolles, CH-1700 Fribourg, Switzerland and Institute of Microbiology, ETH Zurich, CH- 8092 Zurich, Switzerland.
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