Human erythropoietic protoporphyria: identification of a mutation at the splice donor site of intron 7 causing exon 7 skipping of the ferrochelatase gene

Nakahashi, Yoshitsugu; Mitsuya, Hiroaki; Kadota, Yoichi; Naitoh, Yasuyuki; Inoue, Katsumi; Yamamoto, Masayuki; Hayashi, Norio; Taketani, Shigeru

doi:10.1093/hmg/2.7.1069

Cited by 32 publications

(10 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The IVS7+1G>A mutation results in skipping of the 99-bp fragment of exon 7 with the loss of amino acids 236-268 in the final protein product. 49 Although the skipping of this exon does not result in a frameshift, the transcript is likely to produce an inactive enzyme presumably as a result of significant structural variations. 50 We predict that the novel 356_362delTTCAAGA deletion results in a frameshift and a truncated protein of 141 amino acids.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of erythropoietic protoporphyria in South Africa

Parker¹,

Corrigall²,

Hift³

et al. 2008

Br J Dermatol

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

Molecular characterization of erythropoietic protoporphyria in South Africa

Parker¹,

Corrigall²,

Hift³

et al. 2008

Br J Dermatol

View full text Add to dashboard Cite

show abstract

“…A genetic defect in human ferrochelatase causes the disease erythropoietic protoporhyria. Exon deletions are common causes of this disease (32)(33)(34)(35)(36). A set of mutants containing individual deletions of exons 3 through 11 was previously constructed to study the underlying biochemical nature of erythropoietic protoporhyria (37).…”

Section: Characterization Of the C196s Mutant Of Humanmentioning

confidence: 99%

Evidence That the Fourth Ligand to the [2Fe-2S] Cluster in Animal Ferrochelatase Is a Cysteine

Sellers

Wang

Johnson

et al. 1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

The terminal enzyme in heme biosynthesis, ferrochelatase (E.C. 4.99.1.1), catalyzes the insertion of iron into protoporphyrin IX. Nuclear-encoded and produced in the cytoplasm, ferrochelatase is proteolytically processed upon translocation into the mitochondrion. In eukaryotes, the mature-length 42,000 Da protein is associated with the inner mitochondrial membrane, with the active site facing the mitochondrial matrix (1). The proposed catalytic mechanism (2) initially involves a metaldependent, enzyme-mediated distortion of the bound porphyrin ring allowing rapid insertion of Fe 2ϩ into the bent porphyrin (3). Distinct from the enzyme's catalytic activity, a labile 2ϩ cluster has been identified in several animal ferrochelatases including human (4), mouse (5), chicken, and frog (6). Although similar to the [2Fe-2S] 2ϩ centers found in plant ferredoxins, the ferrochelatase iron-sulfur cluster is more labile with enhanced sensitivity to degradation by nitric oxide (7). That the cluster plays no direct role part in catalysis is evidenced by the observation that bacterial, plant, and yeast ferrochelatases do not contain the metal center, and by studies showing that the redox state of the cluster is inconsequential to enzyme activity (4). However, when the cluster is disassembled, enzyme activity is lost (4, 7) and the protein readily precipitates. Hence the cluster appears to play a crucial role in maintaining protein structure in animal ferrochelatases and coupled with the sensitivity to nitric oxide, this has lead us to postulate that the cluster may serve as a regulatory NO 1 -sensor as part of an immune response (7).Site-directed mutagenesis of the five conserved cysteines closest to the COOH terminus of recombinant human ferrochelatase, identified Cys-403, Cys-406, and Cys-411 as ligands to the [2Fe-2S] cluster (8). Additionally, at this time it was proposed that certain spectroscopic anomalies and the unusual lability of the cluster may result from one noncysteinyl oxygenic ligand. The objective of the present investigation was to identify the fourth cluster ligand via additional mutagenesis experiments and by cloning, expression, and characterization of cluster-containing ferrochelatases from more distantly related organisms. We report here the first characterization of ferrochelatase from Drosophila melanogaster and, together with the new mutagenesis results with the human enzyme, these new data support the assignment of the cysteine located at position 196 in the human ferrochelatase as the fourth cluster ligand. Tracing the biological evolution of the ironsulfur cluster in ferrochelatase also provides further insight concerning the specific role of this metal center. EXPERIMENTAL PROCEDURESStrains and Cell Culture-Escherichia coli strain JM109 was used to express recombinant human ferrochelatase, mutant human ferrochelatase, and recombinant Drosophila ferrochelatase as described elsewhere (9). For the mutagenesis procedure described below, E. coli strain BMH mut-s was used to amplify the mutant plasmid. E...

show abstract

“…The molecular cloning and sequence analysis of human FECH cDNA was reported in 1990; 2 subsequently, as many as 90 different mutations affecting the coding regions or splicing junctions have been identified in EPP patients. 3 Our search of the Englishlanguage published work found six Japanese EPP patients in whom genetic analysis of the FECH gene was performed; [4][5][6][7][8][9] another four patients were reported in the Japanese published work or in conference abstracts. An intronic single nucleotide polymorphism (SNP) of the FECH gene, IVS3-48T/C, has been shown to modulate the use of a constitutive aberrant acceptor splice site.…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis of the ferrochelatase gene in eight Japanese patients from seven families with erythropoietic protoporphyria

Saruwatari

Ueki

Yotsumoto

et al. 2006

The Journal of Dermatology

View full text Add to dashboard Cite

A decrease in the activity of ferrochelatase (FECH; EC 4.99.1.1), the terminal enzyme of the heme biosynthetic pathway, results in erythropoietic protoporphyria (EPP; MIM 177000). We analyzed the FECHgene in eight Japanese EPP patients from seven non-consanguineous families and found two distinct genomic DNA abnormalities. In six patients from five families, there was a G-to-A point-mutation at the first position of the intron 9 donor site; it resulted in aberrant splicing and skipping of exon 9 in FECH mRNA. In one patient, we found an A-to-G point-mutation 4 bases from the 3" terminus of intron 4 that led to the in-frame insertion of 3 bases in mRNA. No allelic anomalies, except for 3 single nucleotide polymorphisms were detected in another patient. We analyzed intron polymorphism at IVS3-48, known to be associated with the phenotypic expression of EPP, in these eight patients and 152 healthy Japanese volunteers. All patients were C/C homozygous for IVS3-48. The allelic frequency of IVS3-48C polymorphism in the healthy Japanese volunteers was 67.8% (103/152).

show abstract

Human erythropoietic protoporphyria: identification of a mutation at the splice donor site of intron 7 causing exon 7 skipping of the ferrochelatase gene

Cited by 32 publications

References 0 publications

Molecular characterization of erythropoietic protoporphyria in South Africa

Molecular characterization of erythropoietic protoporphyria in South Africa

Evidence That the Fourth Ligand to the [2Fe-2S] Cluster in Animal Ferrochelatase Is a Cysteine

Genetic analysis of the ferrochelatase gene in eight Japanese patients from seven families with erythropoietic protoporphyria

Contact Info

Product

Resources

About