The cargo receptor NCOA4 mediates autophagic ferritin degradation. Here we show that NCOA4 deficiency in a knockout mouse model causes iron accumulation in the liver and spleen, increased levels of transferrin saturation, serum ferritin, and liver hepcidin, and decreased levels of duodenal ferroportin. Despite signs of iron overload, NCOA4-null mice had mild microcytic hypochromic anemia. Under an iron-deprived diet (2–3 mg/kg), mice failed to release iron from ferritin storage and developed severe microcytic hypochromic anemia and ineffective erythropoiesis associated with increased erythropoietin levels. When fed an iron-enriched diet (2 g/kg), mice died prematurely and showed signs of liver damage. Ferritin accumulated in primary embryonic fibroblasts from NCOA4-null mice consequent to impaired autophagic targeting. Adoptive expression of the NCOA4 COOH terminus (aa 239–614) restored this function. In conclusion, NCOA4 prevents iron accumulation and ensures efficient erythropoiesis, playing a central role in balancing iron levels in vivo. Display omitted •Genetic disruption of NCOA4 causes ferritin accumulation in tissues•NCOA4 deficiency blocks iron mobilization from ferritin storage and induces anemia•NCOA4-null mice are hypersensitive to iron overload NCOA4 is crucial for autophagic ferritin degradation. Bellelli et al. find that genetic disruption of NCOA4 causes ferritin accumulation in tissues and defective iron mobilization from storage. NCOA4-null mice display impaired erythropoiesis associated with microcytic and hypochromic anemia, which is more pronounced in mice fed an iron-depleted diet.