title: Analysis of SLC40A1 gene at the mRNA level reveals rapidly the causative mutations in patients with hereditary hemochromatosis type IV.
abstract: Mutations in the SLC40A1 gene result in a dominant genetic disorder [ferroportin disease; hereditary hemochromatosis type (HH) IV], characterized by iron overload with two different clinical manifestations, normal transferrin saturation with macrophage iron accumulation (the most prevalent type) or high transferrin saturation with hepatocyte iron accumulation (classical hemochromatosis phenotype). In previous studies, the mutational analysis of SLC40A1 gene has been performed at the genomic DNA level by PCR amplification and direct sequencing of all coding regions and flanking intron-exon boundaries (usually in 9 PCR reactions). In this study, we analyzed the SLC40A1 gene at the mRNA level, in two RT-PCR reactions, followed by direct sequencing and/or NIRCA (non-isotopic RNase cleavage assay). This protocol turned out to be rapid, sensitive and reliable, facilitating the detection of the SLC40A1 gene mutations in two patients with hyperferritinemia, normal transferrin saturation and iron accumulation predominantly in macrophages and Kupffer cells. The first one displayed the well-described alteration V162 Delta and the second a novel mutation (R178G) that was further detected in two relatives in a pedigree analysis. The proposed procedure would facilitate the wide-range molecular analysis of the SLC40A1 gene, contributing to better understanding the pathogenesis of the ferroportin disease.
authors.affiliation: Department of Immunology and Histocompatibility, University of Thessaly Medical School, University Hospital of Larissa, 41110 Larissa, Greece. maspel@med.uth.gr
authors.forename: Matthaios
authors.initials: M
authors.lastname: Speletas
authors.forename: Anna
authors.initials: A
authors.lastname: Kioumi
authors.forename: Gedeon
authors.initials: G
authors.lastname: Loules
authors.forename: Prodromos
authors.initials: P
authors.lastname: Hytiroglou
authors.forename: John
authors.initials: J
authors.lastname: Tsitouridis
authors.forename: John
authors.initials: J
authors.lastname: Christakis
authors.forename: Anastasios E
authors.initials: AE
authors.lastname: Germenis
mesh_terms: D000328:Adult; D000369:Aged, 80 and over; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D005293:Ferritins / Q000097:blood; D005838:Genotype; D006432:Hemochromatosis / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000097:blood; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000098742:Ferroportin
chemical_list: D027682:Cation Transport Proteins; D012333:RNA, Messenger; D000098742:Ferroportin; D005293:Ferritins; D007501:Iron