The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans |
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| Authors: | Zvonimir Marelja Mita Mullick Chowdhury Carsten Dosche Carsten Hille Otto Baumann Hans-Gerd L?hmannsr?ben Silke Leimkühler |
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| Affiliation: | 1. Department of Molecular Enzymology, Institute of Biochemistry, University of Potsdam, Potsdam, Germany.; 2. Department of Physical Chemistry, Institute of Chemistry, University of Potsdam, Potsdam, Germany.; 3. Department of Animal Physiology, Institute of Biochemistry, University of Potsdam, Potsdam, Germany.; Queen Mary University of London, United Kingdom, |
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| Abstract: | In humans, the L-cysteine desulfurase NFS1 plays a crucial role in the mitochondrial iron-sulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic tRNAs. We have previously demonstrated that purified NFS1 is able to transfer sulfur to the C-terminal domain of MOCS3, a cytosolic protein involved in molybdenum cofactor biosynthesis and tRNA thiolation. However, no direct evidence existed so far for the interaction of NFS1 and MOCS3 in the cytosol of human cells. Here, we present direct data to show the interaction of NFS1 and MOCS3 in the cytosol of human cells using Förster resonance energy transfer and a split-EGFP system. The colocalization of NFS1 and MOCS3 in the cytosol was confirmed by immunodetection of fractionated cells and localization studies using confocal fluorescence microscopy. Purified NFS1 was used to reconstitute the lacking molybdoenzyme activity of the Neurospora crassa nit-1 mutant, giving additional evidence that NFS1 is the sulfur donor for Moco biosynthesis in eukaryotes in general. |
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