Abnormal mitoferrin-1 expression in patients with erythropoietic protoporphyria.

[erythropoietic protoporphyria]

Most patients with erythropoietic protoporphyria have deficient ferrochelatase (FECH ) activity due to changes in FECH DNA . We evaluated seven patients with erythropoietic protoporphyria phenotype in whom abnormalities of FECH DNA were not found by conventional analysis . The major focus was mitoferrin-1 (MFRN 1 ), the mitochondrial transporter of Fe used for heme formation by FECH and for 2 Fe 2 S cluster synthesis , which is critical to FECH activity /stability .Four patients had a deletion in ALAS 2 that causes enzyme gain-of-function , resulting in increased formation of protoporphyrin ; one had a heterozygous major deletion in FECH DNA . All had an abnormal transcript of MFRN 1 in messenger RNA extracted from blood leukocytes and /or liver tissue . The abnormal transcript contained an insert of intron 2 that had a stop codon . The consequences of abnormal MFRN 1 expression were examined using zebrafish and yeast MFRN-deficient strains and cultured lymphoblasts from the patients .Abnormal human MFRN 1 complementary DNA showed loss -of-function in zebrafish and yeast mutants , whereas normal human MFRN 1 complementary DNA rescued both . Using cultured lymphoblasts , quantitative reverse transcription polymerase chain reaction showed increased formation of abnormal transcript that was accompanied by decreased formation of normal transcript and reduced FECH activity in patients compared to normal lines . A positive correlation coefficient (0 .75 ) was found between FECH activity and normal MFRN 1 messenger RNA in lymphoblasts . However , no obvious cause for increased formation of abnormal transcript was identified in MFRN 1 exons and splice junctions .Abnormal MFRN 1 expression can contribute to erythropoietic protoporphyria phenotype in some patients , probably by causing a reduction in FECH activity .