Abstract
Electron transfer flavoprotein (ETF) is a heterodimeric enzyme composed of an α-subunit and a β-subunit and contains a single equivalent of FAD per dimer. ETF deficiency can be demonstrated in individuals affected by a severe metabolic disorder, glutaric acidemia type II (GAII). In this study, we have investigated for the first time the molecular basis of β-ETF deficiency in three GAII patients: two Japanese brothers, P411 and P412, and a third unrelated patient, P485. Molecular analysis of the β-ETF gene in P411 and P412 demonstrated that both these patients are compound heterozygotes. One allele is carrying a G to A transition at nucleotide 518, causing a missense mutation at codon 164. This point mutation is maternally derived and is not detected in 42 unrelated controls. The other allele carries a G to C transversion at the first nucleotide of the intron donor site, downstream of an exon that is skipped during the splicing event. The sequence analysis of the β-ETF coding sequence in P485 showed only a C to T transition at nucleotide 488 that causes a Thr 154 to Met substitution and the elimination of a Hgal restriction site. Hgal restriction analysis on 63 unrelated controls genomic DNA demonstrated that the C488T transition identifies a polymorphic site. Finally, transfection of wild-type β-ETF cDNA into P411 fibroblasts suggests that wild-type β-ETF cDNA complements the genetic defect and restores the β-oxidation flux to normal levels.
| Original language | English |
|---|---|
| Pages (from-to) | 429-435 |
| Number of pages | 7 |
| Journal | Human Molecular Genetics |
| Volume | 3 |
| Issue number | 3 |
| Publication status | Published - Mar 1994 |
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ASJC Scopus subject areas
- Genetics
- Statistics, Probability and Uncertainty
- Applied Mathematics
- Public Health, Environmental and Occupational Health
- Molecular Biology
- Genetics(clinical)
Cite this
Mutations and polymorphisms of the gene encoding the β-subunit of the electron transfer flavoprotein in three patients with glutaric acidemia type II. / Colombo, Irma; Finocchiaro, Gaetano; Garavaglia, Barbara; Garbuglio, Nicoletta; Yamaguchi, S.; Frerman, F. E.; Berra, Bruno; DiDonato, Stefano.
In: Human Molecular Genetics, Vol. 3, No. 3, 03.1994, p. 429-435.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mutations and polymorphisms of the gene encoding the β-subunit of the electron transfer flavoprotein in three patients with glutaric acidemia type II
AU - Colombo, Irma
AU - Finocchiaro, Gaetano
AU - Garavaglia, Barbara
AU - Garbuglio, Nicoletta
AU - Yamaguchi, S.
AU - Frerman, F. E.
AU - Berra, Bruno
AU - DiDonato, Stefano
PY - 1994/3
Y1 - 1994/3
N2 - Electron transfer flavoprotein (ETF) is a heterodimeric enzyme composed of an α-subunit and a β-subunit and contains a single equivalent of FAD per dimer. ETF deficiency can be demonstrated in individuals affected by a severe metabolic disorder, glutaric acidemia type II (GAII). In this study, we have investigated for the first time the molecular basis of β-ETF deficiency in three GAII patients: two Japanese brothers, P411 and P412, and a third unrelated patient, P485. Molecular analysis of the β-ETF gene in P411 and P412 demonstrated that both these patients are compound heterozygotes. One allele is carrying a G to A transition at nucleotide 518, causing a missense mutation at codon 164. This point mutation is maternally derived and is not detected in 42 unrelated controls. The other allele carries a G to C transversion at the first nucleotide of the intron donor site, downstream of an exon that is skipped during the splicing event. The sequence analysis of the β-ETF coding sequence in P485 showed only a C to T transition at nucleotide 488 that causes a Thr 154 to Met substitution and the elimination of a Hgal restriction site. Hgal restriction analysis on 63 unrelated controls genomic DNA demonstrated that the C488T transition identifies a polymorphic site. Finally, transfection of wild-type β-ETF cDNA into P411 fibroblasts suggests that wild-type β-ETF cDNA complements the genetic defect and restores the β-oxidation flux to normal levels.
AB - Electron transfer flavoprotein (ETF) is a heterodimeric enzyme composed of an α-subunit and a β-subunit and contains a single equivalent of FAD per dimer. ETF deficiency can be demonstrated in individuals affected by a severe metabolic disorder, glutaric acidemia type II (GAII). In this study, we have investigated for the first time the molecular basis of β-ETF deficiency in three GAII patients: two Japanese brothers, P411 and P412, and a third unrelated patient, P485. Molecular analysis of the β-ETF gene in P411 and P412 demonstrated that both these patients are compound heterozygotes. One allele is carrying a G to A transition at nucleotide 518, causing a missense mutation at codon 164. This point mutation is maternally derived and is not detected in 42 unrelated controls. The other allele carries a G to C transversion at the first nucleotide of the intron donor site, downstream of an exon that is skipped during the splicing event. The sequence analysis of the β-ETF coding sequence in P485 showed only a C to T transition at nucleotide 488 that causes a Thr 154 to Met substitution and the elimination of a Hgal restriction site. Hgal restriction analysis on 63 unrelated controls genomic DNA demonstrated that the C488T transition identifies a polymorphic site. Finally, transfection of wild-type β-ETF cDNA into P411 fibroblasts suggests that wild-type β-ETF cDNA complements the genetic defect and restores the β-oxidation flux to normal levels.
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M3 - Article
C2 - 7912128
AN - SCOPUS:0028295653
VL - 3
SP - 429
EP - 435
JO - Human Molecular Genetics
JF - Human Molecular Genetics
SN - 0964-6906
IS - 3
ER -