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The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans

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 c...

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Autores principales: Marelja, Zvonimir, Mullick Chowdhury, Mita, Dosche, Carsten, Hille, Carsten, Baumann, Otto, Löhmannsröben, Hans-Gerd, Leimkühler, Silke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3625234/
https://www.ncbi.nlm.nih.gov/pubmed/23593335
http://dx.doi.org/10.1371/journal.pone.0060869
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author Marelja, Zvonimir
Mullick Chowdhury, Mita
Dosche, Carsten
Hille, Carsten
Baumann, Otto
Löhmannsröben, Hans-Gerd
Leimkühler, Silke
author_facet Marelja, Zvonimir
Mullick Chowdhury, Mita
Dosche, Carsten
Hille, Carsten
Baumann, Otto
Löhmannsröben, Hans-Gerd
Leimkühler, Silke
author_sort Marelja, Zvonimir
collection PubMed
description 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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spelling pubmed-36252342013-04-16 The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans Marelja, Zvonimir Mullick Chowdhury, Mita Dosche, Carsten Hille, Carsten Baumann, Otto Löhmannsröben, Hans-Gerd Leimkühler, Silke PLoS One Research Article 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. Public Library of Science 2013-04-12 /pmc/articles/PMC3625234/ /pubmed/23593335 http://dx.doi.org/10.1371/journal.pone.0060869 Text en © 2013 Marelja et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Marelja, Zvonimir
Mullick Chowdhury, Mita
Dosche, Carsten
Hille, Carsten
Baumann, Otto
Löhmannsröben, Hans-Gerd
Leimkühler, Silke
The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title_full The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title_fullStr The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title_full_unstemmed The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title_short The L-Cysteine Desulfurase NFS1 Is Localized in the Cytosol where it Provides the Sulfur for Molybdenum Cofactor Biosynthesis in Humans
title_sort l-cysteine desulfurase nfs1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3625234/
https://www.ncbi.nlm.nih.gov/pubmed/23593335
http://dx.doi.org/10.1371/journal.pone.0060869
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