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The substrate and function of mitochondrial ABC transporters involved in Fe-S metabolism

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ABC transporters of the mitochondria (ATM) are required for cytosolic Fe-S cluster assembly. They are located in the inner membrane of the mitochondria and mediate ATP -driven transport of a so-far unknown compound from the mitochondria into the cytosol. Previous studies in the model plant Arabidopsis suggested that the transported compound contains sulphur (but not iron) in an unknown chemical form (1). Additionally, a role of GSH in yeast Atm1 function was shown (2).

The aim of our study is to characterize the substrate specificity of the plant AtATM3 and its orthologue yeast ScAtm1. We analyzed sulphur and thiol containing compounds in isolated mitochondria using High Pressure Liquid Chromatography and observed changes of the ratios of GSH to other thiols in atm mutants in both yeast and Arabidopsis. Arabidopsis atm3 mutants are also sensitive to buthionine sulfoximine (BSO), an inhibitor of the first step of glutathione synthesis, and as a result display reduced root growth. These data indicate that GSH is important in ATM function – as previously suggested for yeast Atm1 (2).

To further investigate the substrate specificity of ATMs we have established an expression and purification system for ATMs in the gram-positive bacterium Lactococcus lactis and are currently screening for potential substrates by monitoring the rate of ATP hydrolysis of purified potein. Future work will include establishment of transport assays using the lactococcal system.

1. Bernard, D. et al. (2009) Plant Physiol. 151: 590-602

2. Sipos, K. et al. (2002) J. Biol. Chem. 277(30): 26944-26949

This talk is part of the Plant Sciences Research Seminars series.

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