title: A deficiency in dolichyl-P-glucose:Glc1Man9GlcNAc2-PP-dolichyl alpha3-glucosyltransferase defines a new subtype of congenital disorders of glycosylation.
abstract: The underlying causes of type I congenital disorders of glycosylation (CDG I) have been shown to be mutations in genes encoding proteins involved in the biosynthesis of the dolichyl-linked oligosaccharide (Glc(3)Man(9)GlcNAc(2)-PP-dolichyl) that is required for protein glycosylation. Here we describe a CDG I patient displaying gastrointestinal problems but no central nervous system deficits. Fibroblasts from this patient accumulate mainly Man(9)GlcNAc(2)-PP-dolichyl, but in the presence of castanospermine, an endoplasmic reticulum glucosidase inhibitor Glc(1)Man(9)GlcNAc(2)-PP-dolichyl predominates, suggesting inefficient addition of the second glucose residue onto lipid-linked oligosaccharide. Northern blot analysis revealed the cells from the patient to possess only 10-20% normal amounts of mRNA encoding the enzyme, dolichyl-P-glucose:Glc(1)Man(9)GlcNAc(2)-PP-dolichyl alpha3-glucosyltransferase (hALG8p), which catalyzes this reaction. Sequencing of hALG8 genomic DNA revealed exon 4 to contain a base deletion in one allele and a base insertion in the other. Both mutations give rise to premature stop codons predicted to generate severely truncated proteins, but because the translation inhibitor emetine was shown to stabilize the hALG8 mRNA from the patient to normal levels, it is likely that both transcripts undergo nonsense-mediated mRNA decay. As the cells from the patient were successfully complemented with wild type hALG8 cDNA, we conclude that these mutations are the underlying cause of this new CDG I subtype that we propose be called CDG Ih.
authors.affiliation: Unité de Glycobiologie et Signalisation Cellulaire, INSERM, U504, Bâtiment INSERM, 16 Avenue Paul Vaillant-Couturier, 94807 Villejuif, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Chantret
authors.forename: Julia
authors.initials: J
authors.lastname: Dancourt
authors.forename: Thierry
authors.initials: T
authors.lastname: Dupré
authors.forename: Christophe
authors.initials: C
authors.lastname: Delenda
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Bucher
authors.forename: Sandrine
authors.initials: S
authors.lastname: Vuillaumier-Barrot
authors.forename: Hélène
authors.initials: H
authors.lastname: Ogier de Baulny
authors.forename: Celine
authors.initials: C
authors.lastname: Peletan
authors.forename: Olivier
authors.initials: O
authors.lastname: Danos
authors.forename: Nathalie
authors.initials: N
authors.lastname: Seta
authors.forename: Geneviève
authors.initials: G
authors.lastname: Durand
authors.forename: Rafael
authors.initials: R
authors.lastname: Oriol
authors.forename: Patrice
authors.initials: P
authors.lastname: Codogno
authors.forename: Stuart E H
authors.initials: SE
authors.lastname: Moore
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002239:Carbohydrate Metabolism, Inborn Errors / Q000175:diagnosis / Q000201:enzymology* / Q000235:genetics; D002478:Cells, Cultured; D002725:Chloroform / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D018388:Codon, Terminator; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008214:Lymphocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009844:Oligosaccharides / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors
chemical_list: D018388:Codon, Terminator; D018076:DNA, Complementary; D008055:Lipids; D009844:Oligosaccharides; D012333:RNA, Messenger; D002725:Chloroform; C473777:ALG8 protein, human; D005964:Glucosyltransferases; C054676:dolichol phosphate glucose-dolichol diphosphate oligosaccharide glucosyltransferase