Nitrogen monoxide-mediated control of ferritin synthesis: implications for macrophage iron homeostasis.
Intracellular iron homeostasis is regulated posttranscriptionally by iron regulatory proteins 1 and 2 (IRP1 and IRP2). In the absence of iron in the labile pool, IRPs bind to specific nucleotide sequences called iron responsive elements (IREs), which are located in the 5' untranslated region of ferritin mRNA and the 3' untranslated region of transferrin receptor mRNA. IRP binding to the IREs suppresses ferritin translation and stabilizes transferrin receptor mRNA, whereas the opposite scenario develops in iron-replete cells. Binding of IRPs to the IREs is also affected by nitrogen monoxide (NO), but there are conflicting reports regarding the effect of NO on ferritin synthesis. In this study, we demonstrated that a short exposure of RAW 264.7 cells (a macrophage cell line) to the NO+ donor, sodium nitroprusside (SNP), resulted in a dramatic increase in ferritin synthesis. The SNP-mediated increase of ferritin synthesis could be blocked by MG132, an inhibitor of proteasome-dependent protein degradation, which also prevented the degradation of IRP2 caused by SNP treatment. Moreover, treatment of RAW 264.7 cells with IFN-gamma and lipopolysaccharide caused IRP2 degradation and stimulated ferritin synthesis, changes that could be prevented by specific inhibitors of inducible nitric oxide synthase. Furthermore, the SNP-mediated increase in ferritin synthesis was associated with a significant enhancement of iron incorporation into ferritin. These observations indicate that NO+-mediated modulation of IRP2 plays an important role in controlling ferritin synthesis and iron metabolism in murine macrophages.
Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital and Department of Physiology, McGill University, 3755 Côte Sainte-Catherine Road, Montreal, QC, Canada H3T 1E2.
Sangwon
S
Kim
Prem
P
Ponka
D000818:Animals; D002460:Cell Line; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D005293:Ferritins / Q000096:biosynthesis*; D006706:Homeostasis; D007371:Interferon-gamma / Q000494:pharmacology; D007501:Iron / Q000378:metabolism*; D035941:Iron Regulatory Protein 1; D035942:Iron Regulatory Protein 2; D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D007700:Kinetics; D007976:Leupeptins / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000378:metabolism; D011994:Recombinant Proteins
D015853:Cysteine Proteinase Inhibitors; D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins; D007976:Leupeptins; D008070:Lipopolysaccharides; D020030:Nitric Oxide Donors; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D009599:Nitroprusside; D009569:Nitric Oxide; D007371:Interferon-gamma; D005293:Ferritins; D007501:Iron; D035941:Iron Regulatory Protein 1; D035942:Iron Regulatory Protein 2; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde