The L-cysteine desulfurase NFS1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans

Zvonimir Marelja; Mita Mullick Chowdhury; Carsten Dosche; Carsten Hille; Otto Baumann; Hans-Gerd Löhmannsröben; Silke Leimkühler

doi:10.1371/journal.pone.0060869

The L-cysteine desulfurase NFS1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans

PLoS One. 2013 Apr 12;8(4):e60869. doi: 10.1371/journal.pone.0060869. Print 2013.

Authors

Zvonimir Marelja¹, Mita Mullick Chowdhury, Carsten Dosche, Carsten Hille, Otto Baumann, Hans-Gerd Löhmannsröben, Silke Leimkühler

Affiliation

¹ Department of Molecular Enzymology, Institute of Biochemistry, University of Potsdam, Potsdam, Germany.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbon-Sulfur Lyases / metabolism*
Coenzymes / biosynthesis*
Cysteine / metabolism*
Cytosol / metabolism*
Fluorescence Resonance Energy Transfer
Green Fluorescent Proteins / metabolism
HeLa Cells
Humans
Metalloproteins / biosynthesis*
Microscopy, Fluorescence
Molybdenum Cofactors
Mutant Proteins / metabolism
Neurospora / enzymology
Nitrate Reductase / metabolism
Nucleotidyltransferases / metabolism
Protein Interaction Mapping
Protein Transport
Pteridines
Recombinant Fusion Proteins / metabolism
Subcellular Fractions / metabolism
Sulfur / metabolism*
Sulfurtransferases / metabolism
Surface Plasmon Resonance

Substances

Coenzymes
Metalloproteins
Molybdenum Cofactors
Mutant Proteins
Pteridines
Recombinant Fusion Proteins
enhanced green fluorescent protein
Green Fluorescent Proteins
Sulfur
molybdenum cofactor
Nitrate Reductase
MOCS3 protein, human
Nucleotidyltransferases
Sulfurtransferases
Carbon-Sulfur Lyases
NFS1 protein, human
Cysteine

Grants and funding

This work was supported by the Studienstiftung des deutschen Volkes and the DFG grant LE1171/5-3 (to S.L.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.