New and old players in the hepcidin pathway

The identification of the liver peptide hepcidin at the beginning of the new millennium opened a new era in our understanding of iron metabolism.[1][1]–[3][2] Hepcidin is the main regulator of intestinal iron absorption and macrophage iron release, thus ultimately of the iron available fo

Furin-mediated release of soluble hemojuvelin: a new link between hypoxia and iron homeostasis

The liver peptide hepcidin regulates iron absorption and recycling. Hemojuvelin (HJV) has a key role in hepcidin regulation, and its inactivation causes severe iron overload both in humans and in mice. Membrane HJV (m-HJV) acts as a coreceptor for bone morphogenetic proteins (BMPs), whereas soluble HJV (s-HJV) may down-regulate hepcidin in a competitive way interfering with BMP signaling. s-HJV is decreased by iron in vitro and increased by iron deficiency in vivo. However, the mechanisms regulating the 2 HJV isoforms remain unclear. Here we show that s-HJV originates from a furin cleavage at position 332–335. s-HJV is reduced in the cleavage mutant R335Q as well as in cells treated with a furin inhibitor, and increased in cells overexpressing exogenous furin, but not in cells overexpressing an inactive furin variant. Furin is up-regulated by iron deficiency and hypoxia in association with the stabilization of HIF-1α. Increased s-HJV in response to HIF-1α occurs during differentiation of murine muscle cells expressing endogenous Hjv. Our data are relevant to the mechanisms that relate iron metabolism to the hypoxic response. The release of s-HJV might be a tissue-specific mechanism, signaling the local iron requests of hypoxic skeletal muscles independently of the oxygen status of the liver

Hemojuvelin N-terminal mutants reach the plasma membrane but do not activate the hepcidin response

Background Hemojuvelin is a glycosylphosphatidylinositol-anchored protein, expressed in liver, skeletal muscle and heart. As a co-receptor of bone morphogenetic protein, membrane hemojuvelin positively modulates the iron regulator hepcidin. Mutations of the gene encoding for hemojuvelin cause juvenile hemochromatosis, characterized by hepcidin deficiency and severe iron overload. We have previously shown that several hemojuvelin variants do not efficiently reach the plasma membrane, whereas a few N-terminal mutants localize to the plasma membrane. Design and Methods We studied hemojuvelin mutants of N-terminus (C80R, S85P, G99V, ΔRGD) and GDPH-consensus site for autoproteolysis (A168D, F170S, D172E) transiently expressed in HeLa cells, using electron microscopy, morphometric analysis and binding assays at different time points. Hepcidin activation by wild-type and mutant forms of hemojuvelin was assessed in Hep3B cells transfected with a hepcidin-promoter luciferase-reporter construct. Results S85P, G99V and ΔRGD were localized to plasma membrane 36 hours after transfection, but less efficiently exported than the wild-type protein at earlier (24–30 hours) times. Morphometric analysis clearly documented delayed export and endoplasmic reticulum retention of G99V. C80R was exported without delay. GDPH variants were partially retained in the endoplasmic reticulum and Golgi apparatus, but showed impaired plasma membrane localization. In the hepcidin promoter assay only wild type hemojuvelin was able to activate hepcidin. Conclusions The delayed export and retention in the endoplasmic reticulum of some N-terminal mutants could contribute to the pathogenesis of juvenile hemochromatosis, reducing a prompt response of bone morphogenetic protein. However, independently of their plasma membrane export, all hemojuvelin mutants tested showed no or minimal hepcidin activation

Two to Tango: Regulation of Mammalian Iron Metabolism

Disruptions in iron homeostasis from both iron deficiency and overload account for some of the most common human diseases. Iron metabolism is balanced by two regulatory systems, one that functions systemically and relies on the hormone hepcidin and the iron exporter ferroportin, and another that predominantly controls cellular iron metabolism through iron-regulatory proteins that bind iron-responsive elements in regulated messenger RNAs. We describe how the two distinct systems function and how they “tango” together in a coordinated manner. We also highlight some of the current questions in mammalian iron metabolism and discuss therapeutic opportunities arising from a better understanding of the underlying biological principles

New TFR2 mutations in young Italian patients with hemochromatosis.

This work describes the identification of two subjects with young-age iron overload carrying new causative mutations in transferrin receptor-2 gene. One was compound heterozygous (Asn411del/Ala444Thr) and the second was homozygous for a mutation affecting RNA splicing (IVS17+5636G>A). Another mutation (His33Asn) and a polymorphism were found in a group of 50 controls

Molecular Basis of Phenylketonuria in Italy

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Construction of a YAC contig covering human chromosome 6p22

A contig covering human chromosome 6p22 that consists of 134 YAC clones aligned based on the presence/absence of 52 DNA markers is presented. This contig overlaps with the 6p23 contig at its telomeric end and with the 6p21.3 contig at its centromeric end. The order of loci within the contig resolves the relative positions of several genetically mapped markers. Among the additional markers used here, there are eight novel PCR assays. The 12 known genes and anonymous ESTs located within the contig establish a first step toward a transcriptional map of this region. The instability of YAC clones observed during this work is also discussed. (C) 1996 Academic Press, Inc

Variation of hemoglobin levels in normal Italian populations from genetic isolates.

Normal hemoglobin levels vary greatly according to genetic and acquired factors. As a consequence there is no general agreement on the definition of anemia in terms of hemoglobin levels. Here we compare the hemoglobin levels of subjects recruited from normal genetically isolated Italian populations whose medical history, life style habits and results of laboratory tests are available. After the exclusion of pathological samples we analyzed the hemoglobin levels of 3,849 subjects (1,661 males and 2,188 females) and evaluated the hemoglobin heritability. Normal subjects of different age groups from a northern Italian isolate have significantly higher hemoglobin levels when compared to matched subjects of southern Italian isolates. The estimated heritability of hemoglobin levels ranges from 0.34 to 0.42 in the different isolates. Our study provides a dataset of hemoglobin levels for normal subjects of different geographical origin and indicate that hemoglobin levels are substantially influenced by heritable components