A 17-bp insertion and a Phe215 → Cys missense mutation in the dihydrolipoyl transacylase (E2) mRNA from a thiamine-responsive maple syrup urine disease patient WG-34

Charles W. Fisher, Kim S. Lau, Carolyn R. Fisher, R. Max Wynn, Rody P. Cox, David T. Chuang

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21 Citations (Scopus)

Abstract

We have amplified the cDNA for the transacylase (E2) subunit of the branched-chain α-ketoacid dehydrogenase (BCKAD) complex from a thiamine-responsive MSUD cell line (WG-34) by the polymerase chain reaction. Sequencing of the amplified WG-34 cDNA showed a 17-bp insertion (AAATACCTTGTTACCAG) apparently resulting from an aberrant splicing of the E2 gene, and a missense (T → G) mutation that changes Phe215 to Cys in the E2 subunit. The existence of these two mutations was confirmed by probing the amplified E2 cDNA or genomic DNA with allele-specific oligonucleotides. The above results support the thesis that the thiamine-responsive MSUD patient (WG-34) is a compound heterozygote at the E2 locus. The implication of the E2 mutations for the thiamine-responsiveness observed in this patient is discussed.

Original languageEnglish (US)
Pages (from-to)804-809
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume174
Issue number2
DOIs
StatePublished - Jan 31 1991

Fingerprint

Maple Syrup Urine Disease
Thiamine
Missense Mutation
Complementary DNA
Messenger RNA
Mutation
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Polymerase chain reaction
Heterozygote
Oligonucleotides
Genes
Alleles
Cells
Cell Line
Polymerase Chain Reaction
DNA
dihydrolipoamide acyltransferase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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title = "A 17-bp insertion and a Phe215 → Cys missense mutation in the dihydrolipoyl transacylase (E2) mRNA from a thiamine-responsive maple syrup urine disease patient WG-34",
abstract = "We have amplified the cDNA for the transacylase (E2) subunit of the branched-chain α-ketoacid dehydrogenase (BCKAD) complex from a thiamine-responsive MSUD cell line (WG-34) by the polymerase chain reaction. Sequencing of the amplified WG-34 cDNA showed a 17-bp insertion (AAATACCTTGTTACCAG) apparently resulting from an aberrant splicing of the E2 gene, and a missense (T → G) mutation that changes Phe215 to Cys in the E2 subunit. The existence of these two mutations was confirmed by probing the amplified E2 cDNA or genomic DNA with allele-specific oligonucleotides. The above results support the thesis that the thiamine-responsive MSUD patient (WG-34) is a compound heterozygote at the E2 locus. The implication of the E2 mutations for the thiamine-responsiveness observed in this patient is discussed.",
author = "Fisher, {Charles W.} and Lau, {Kim S.} and Fisher, {Carolyn R.} and Wynn, {R. Max} and Cox, {Rody P.} and Chuang, {David T.}",
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T1 - A 17-bp insertion and a Phe215 → Cys missense mutation in the dihydrolipoyl transacylase (E2) mRNA from a thiamine-responsive maple syrup urine disease patient WG-34

AU - Fisher, Charles W.

AU - Lau, Kim S.

AU - Fisher, Carolyn R.

AU - Wynn, R. Max

AU - Cox, Rody P.

AU - Chuang, David T.

PY - 1991/1/31

Y1 - 1991/1/31

N2 - We have amplified the cDNA for the transacylase (E2) subunit of the branched-chain α-ketoacid dehydrogenase (BCKAD) complex from a thiamine-responsive MSUD cell line (WG-34) by the polymerase chain reaction. Sequencing of the amplified WG-34 cDNA showed a 17-bp insertion (AAATACCTTGTTACCAG) apparently resulting from an aberrant splicing of the E2 gene, and a missense (T → G) mutation that changes Phe215 to Cys in the E2 subunit. The existence of these two mutations was confirmed by probing the amplified E2 cDNA or genomic DNA with allele-specific oligonucleotides. The above results support the thesis that the thiamine-responsive MSUD patient (WG-34) is a compound heterozygote at the E2 locus. The implication of the E2 mutations for the thiamine-responsiveness observed in this patient is discussed.

AB - We have amplified the cDNA for the transacylase (E2) subunit of the branched-chain α-ketoacid dehydrogenase (BCKAD) complex from a thiamine-responsive MSUD cell line (WG-34) by the polymerase chain reaction. Sequencing of the amplified WG-34 cDNA showed a 17-bp insertion (AAATACCTTGTTACCAG) apparently resulting from an aberrant splicing of the E2 gene, and a missense (T → G) mutation that changes Phe215 to Cys in the E2 subunit. The existence of these two mutations was confirmed by probing the amplified E2 cDNA or genomic DNA with allele-specific oligonucleotides. The above results support the thesis that the thiamine-responsive MSUD patient (WG-34) is a compound heterozygote at the E2 locus. The implication of the E2 mutations for the thiamine-responsiveness observed in this patient is discussed.

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