Abstract
Iron-sulfur (Fe-S) clusters are universal cofactors embedded in proteins that support numerous fundamental biological processes such as photosynthesis and respiration (1). Assembly and delivery of Fe-S centres to client proteins take place within specialised Fe-S assembly machineries involving numerous proteins and cofactors, some of which are still poorly characterised in plants. Considering that more than 200 key client proteins in Arabidopsis require Fe-S clusters to perform their function (2), we are developing research on BOLA proteins known in few model organisms as interactors of two essential classes of Fe-S shuttle proteins within Fe-S assembly machineries (3-5), but their importance and exact role in plants remain to be assessed. Focusing on Arabidopsis thaliana BOLA paralogs, our recent work showed that the cumulative depletion of the plastidial/mitochondrial BOLA1 and BOLA4 proteins leads to a severe dwarf phenotype with early primary root arrest, suggesting important role of these proteins for plant development (6). To go further in understanding the function of these proteins, we describe here selected strategies based on the use of bola1bola4/proBOLA::gBOLA-GFP transgenic lines to unravel their interactomes with a highlight on BOLA4, with the aim to position its function within the mitochondrial and platidial Fe-S assembly machineries.(1) Balk and Schaedler (2014) Iron Cofactor Assembly in Plants. Annual Review of Plant Biology 65: 125-153.(2) Przybyla-Toscano et al. (2021) Gene atlas of iron-containing proteins in Arabidopsis thaliana. The plant journal 106: 258-274.(3) Pedroletti et al. (2023) Journal of Experimental Botany 74: 3328–3344.(4) Azam et al. (2020a) The Arabidopsis Mitochondrial Glutaredoxin GRXS15 Provides [2Fe-2S] Clusters for ISCA- Mediated [4Fe-4S] Cluster Maturation. International Journal of Molecular Sciences 21(23): 9237.(5) Azam et al. (2020b) [4Fe-4S] cluster trafficking mediated by Arabidopsis mitochondrial ISCA and NFU proteins. Journal of Biological Chemistry 295(52): 18367-18378.(6) Taupin-Broggini*, Rodriguez-Euzet* et al. (2024), in preparation.