A connection between two ancient and essential cellular processes, iron-sulfur protein biogenesis and fatty acid...

Iron-sulfur [Fe-S] clusters are ubiquitous cofactors of a wide array of structurally and functionally diverse proteins. Acyl carrier protein (ACP) is the universal factor required for fatty acid (FA) synthesis. In this study, in E. coli, we demonstrated that [Fe-S] and FA biosynthesis pathways are coordinated processes, driven by a physical interaction between ACP and the ISC [Fe-S] biogenesis machinery. Using bacterial two-hybrid assays, co-purification, and biochemical analyses, we demonstrated a molecular interaction between ACP and IscS, the ISC machinery cysteine desulfurase that provides sulfur for [Fe-S] cluster formation. Structural modeling and directed mutagenesis pinpointed the ACP-binding site in a region of IscS shared for interactions with other components of the ISC [Fe-S] biogenesis system, mainly Fdx and CyaY. At the cellular level, ACP depletion was found to disrupt ISC-dependent [Fe-S] cluster biogenesis, diminishing the activity of key [Fe-S]-dependent regulators (IscR, FNR, and NsrR) and enzymes (aconitase and biotin synthase). Moreover, FASII inhibition by triclosan, which perturbs ACP species homeostasis, also compromised the assembly of the [Fe-S]-dependent FNR regulator. Our findings underscore a functional link between [Fe-S] cluster biogenesis and fatty acid metabolism, with far-reaching unexplored intricacies of metabolic coordination and cellular homeostasis. Comparison with eukaryotic systems highlights a strong evolutionary driving force toward a link between [Fe-S] cluster and fatty acid biosynthesis in all living systems.IMPORTANCECellular functions rely on interconnected metabolic pathways; however, many regulatory links remain unexplored. Iron-sulfur [Fe-S] clusters are cofactors of proteins driving fundamental cellular processes, from respiration to gene regulation. Our study uncovers a direct connection between [Fe-S] cluster biogenesis and fatty acid biosynthesis. We demonstrate the molecular connection between these two essential cellular processes to lie within the interaction between the acyl carrier protein (ACP), a shuttle of fatty acid biosynthetic intermediates, and IscS, the source of sulfur for [Fe-S] cluster assembly. Intriguingly, similar interactions between ACP and [Fe-S] building cysteine desulfurase have been observed in yeast and human models; however, these mechanisms rest on different molecular determinants. This points out the existence of a strong evolutive driving force toward establishing a link between [Fe-S] cluster and fatty acid biosynthesis in all living systems, with far-reaching implications for metabolic coordination and cellular homeostasis.