PMID- 10477430
OWN - NLM
STAT- MEDLINE
DCOM- 19991012
LR  - 20091119
IS  - 1059-7794 (Print)
IS  - 1059-7794 (Linking)
VI  - 14
IP  - 3
DP  - 1999
TI  - Screening for mutations in the uroporphyrinogen decarboxylase gene using
      denaturing gradient gel electrophoresis. Identification and characterization of
      six novel mutations associated with familial PCT.
PG  - 222-32
AB  - The two porphyrias, familial porphyria cutanea tarda (fPCT) and
      hepatoerythropoietic porphyria (HEP), are associated with mutations in the gene
      encoding the enzyme uroporphyrinogen decarboxylase (UROD). Several mutations,
      most of which are private, have been identified in HEP and fPCT patients,
      confirming the heterogeneity of the underlying genetic defects of these diseases.
      We have established a denaturing gradient gel electrophoresis (DGGE) assay for
      mutation detection in the UROD gene, enabling the simultaneous screening for
      known and unknown mutations. The established assay has proved able to detect the 
      underlying UROD mutation in 10 previously characterized DNA samples as well as a 
      new mutation in each of six previously unexamined PCT patients. The six novel
      UROD mutations comprise three missense mutations (M01T, F229L, and M324T), two
      splice mutations (IVS3-2A-->T and IVS5-2A-->G) leading to exon skipping, and a
      2-bp deletion (415-416delTA) resulting in a frameshift and the introduction of a 
      premature stop codon. Heterologous expression and enzymatic studies of the mutant
      proteins demonstrate that the three mutations leading to shortening or truncation
      of the UROD protein have no residual catalytic activity, whereas the two missense
      mutants retained some residual activity. Furthermore, the missense mutants
      exhibited a considerable increase in thermolability. The six new mutations bring 
      to a total of 29 the number of disease-related mutations in the UROD gene. The
      DGGE assay presented greatly improves the genetic diagnosis of fPCT and HEP,
      thereby facilitating the detection of familial UROD deficient patients as well as
      the discrimination between familial and sporadic PCT cases.
CI  - Copyright 1999 Wiley-Liss, Inc.
FAU - Christiansen, L
AU  - Christiansen L
AD  - Department of Clinical Biochemistry and Clinical Genetics, Odense University
      Hospital, Odense, Denmark. lec@imbmed.ou.dk
FAU - Ged, C
AU  - Ged C
FAU - Hombrados, I
AU  - Hombrados I
FAU - Brons-Poulsen, J
AU  - Brons-Poulsen J
FAU - Fontanellas, A
AU  - Fontanellas A
FAU - de Verneuil, H
AU  - de Verneuil H
FAU - Horder, M
AU  - Horder M
FAU - Petersen, N E
AU  - Petersen NE
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Hum Mutat
JT  - Human mutation
JID - 9215429
RN  - 0 (Recombinant Proteins)
RN  - EC 4.1.1.37 (Uroporphyrinogen Decarboxylase)
SB  - IM
MH  - Alleles
MH  - DNA Mutational Analysis
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Enzyme Stability
MH  - Exons/genetics
MH  - Gene Expression
MH  - Genetic Testing/*methods
MH  - Heterozygote
MH  - Homozygote
MH  - Humans
MH  - Mutagenesis, Site-Directed
MH  - *Mutation, Missense
MH  - Porphyria Cutanea Tarda/*enzymology
MH  - RNA Splicing/genetics
MH  - Recombinant Proteins/genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sequence Analysis, DNA
MH  - Temperature
MH  - Uroporphyrinogen Decarboxylase/*genetics
EDAT- 1999/09/08 00:00
MHDA- 1999/09/08 00:01
CRDT- 1999/09/08 00:00
PHST- 1999/09/08 00:00 [pubmed]
PHST- 1999/09/08 00:01 [medline]
PHST- 1999/09/08 00:00 [entrez]
AID - 10.1002/(SICI)1098-1004(1999)14:3<222::AID-HUMU5>3.0.CO;2-V [pii]
AID - 10.1002/(SICI)1098-1004(1999)14:3<222::AID-HUMU5>3.0.CO;2-V [doi]
PST - ppublish
SO  - Hum Mutat. 1999;14(3):222-32. doi:
      10.1002/(SICI)1098-1004(1999)14:3<222::AID-HUMU5>3.0.CO;2-V.