GM20941
LCL from B-Lymphocyte
Description:
CONGENITAL DISORDER OF GLYCOSYLATION, TYPE Ij
DOLICHYL-PHOSPHATE N-ACETYLGLUCOSAMINE PHOSPHOTRANSFERASE; DPAGT1
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Repository
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NIGMS Human Genetic Cell Repository
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| Subcollection |
Heritable Diseases |
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Biopsy Source
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Peripheral vein
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Cell Type
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B-Lymphocyte
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Tissue Type
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Blood
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Transformant
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Epstein-Barr Virus
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Sample Source
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LCL from B-Lymphocyte
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Relation to Proband
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proband
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Confirmation
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Molecular characterization before cell line submission to CCR
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Species
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Homo sapiens
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Common Name
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Human
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Remarks
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| alanine transaminase |
According to the submitter biochemical test results for this subject showed decreased enzyme activity. EC Number: 2.6.1.2; <10% activity. |
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| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin confirmed by LINE assay |
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| phosphomannomutase |
According to the submitter biochemical test results for this subject showed normal enzyme activity. EC Number: 5.4.2.8 |
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| Gene |
DPAGT1 |
| Chromosomal Location |
11q23.3 |
| Allelic Variant 1 |
191350.0001; CONGENITAL DISORDER OF GLYCOSYLATION, TYPE Ij |
| Identified Mutation |
TYR170CYS; In a patient with CDG Ij (608093), Wu et al. (Hum Mutat 22:144-150, 2003) identified reduced DPAGT1 enzymatic activity; sequencing of genomic DNA and cDNAs of the DPAGT1 gene identified, in the paternal allele, a 660A-G transition in exon 5, resulting in a tyr170-to-cys (Y170C) mutation. Although no mutation was identified in the maternal allele, it produced only 12% of the normal amount of mature mRNA; the remainder showed a complex exon skipping pattern that shifted the reading frame and resulted in a truncated nonfunctional protein. |
| Remarks |
Clinically affected; developed infantile spasms at age 4 months within 72 hours of receiving DPT immunization and diagnosis confirmed by hypsarrhythmia on EEG and treated with ACTH; mental retardation; microcephaly; arched palate; micrognathia; exotropia; fifth finger clinodactyly; single flexion creases; skin dimples on upper thighs; severe hypotonia; medically intractable seizures; minimal linguistic ability at age 6 years; normal liver and renal function; normal hematologic parameters; normal karyotype; normal brain MRI at age 6 months; PET scan showed multifocal areas of decreased metabolic activity; abnormal isoelectric focusing pattern of serum transferrin; reduced amount of normal LLO (lipid-linked oligosaccharide); assay for phosphomannomutase (CDG-Ia) and phosphomannose isomerase (CDG-Ib) were normal; GPT activity in microsomes in fibroblasts was less than 10% of the GPT activity in control fibroblasts when assayed either in the presence or absence of exogenous dolichol phosphate; donor subject has one allele with an A>G transition at nucleotide 660 in exon 5 of the DPAGT1 gene [660A>G] resulting in a substitution of cysteine for tyrosine at codon 170 [Tyr170Cys (Y170C)] and the other allele (maternal) produces alternative forms of DPAGT1 in which multiple exons are skipped; the reproducible alternative splicing was only observed in the donor subject and maternal cells; sequencing of entire gene with the exception of intron 5 did not reveal a mutation in the maternal allele; cells from donor subject produce only 12% normal DPAGT1 message from the maternal allele.
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| Wu X, Rush JS, Karaoglu D, Krasnewich D, Lubinsky MS, Waechter CJ, Gilmore R, Freeze HH, Deficiency of UDP-GlcNAc:Dolichol Phosphate N-Acetylglucosamine-1 Phosphate Transferase (DPAGT1) causes a novel congenital disorder of Glycosylation Type Ij Human mutation22:144-50 2003 |
| PubMed ID: 12872255 |
| Split Ratio |
1:4 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Medium |
Roswell Park Memorial Institute Medium 1640 with 2mM L-glutamine or equivalent |
| Serum |
15% fetal bovine serum Not Inactivated |
| Supplement |
- |
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