Thermophilic fungus uses anthraquinones to modulate ferrous excretion, sterol-mediated endocytosis, and iron storage in response to cold stress

To date, there are no real physiological mechanisms for iron excretion in eukaryote, and no physiological “actuator” that can control all the three fundamental biologic processes of absorption, storage, and excretion. Here, we observed that the accumulation of anthraquinones by Thermomyces dupontii under cold stress can achieve this process. Through mutation analysis, we found that mutant ΔAn deficiency in anthraquinones accumulated ferrous and total free iron due to adopting a rare lifestyle with no endocytosis but accumulation of membrane-derived vesicles. Anthraquinone complement indicated that the vesicles in ΔAn could coat the extrinsic anthraquinone-induced granules to prevent contact with the fungal interiors. Detailed chemical investigation on ΔAn led to characterization of a rare oxygen-free ergosterene with unstable nature in air as the major membrane steroid in ΔAn, suggesting hypoxia inner in ΔAn cells, consistent with dramatically low oxygen-consuming rates in ΔAn. A series of physiological and metabolic analyses indicated anthraquinones were involved in exporting ferrous and promoting formation of oxygen-containing metabolites, including ergosterols for endocytosis and iron chelators for iron storage. Moreover, we found that both the anticancer agent mitoxantrone with well-known-cardiotoxicity side effect and the major terpenoid-derived polycyclic aromatics from Danshen for treating cardiovascular disease showed potent ferrous transporting capabilities in human cancer cells. Our findings provide a novel insight into the underlying mechanisms of polycyclic aromatics in nature and pharmacology, and offer a new strategy for developing potential therapeutics and agents for membrane transport, iron homestasis, and anticold.