Functional characterization of the eukaryotic cysteine desulfurase Nfs1p from Saccharomyces cerevisiae

Ulrich Mühlenhoff, Janneke Balk, Nadine Richhardt, Jens T. Kaiser, Katalin Sipos, G. Kispál, Roland Lill

Research output: Contribution to journalArticle

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Abstract

Previous studies have indicated that the essential protein Nfs1 performs a crucial role in cellular iron-sulfur (Fe/S) protein maturation. The protein is located predominantly in mitochondria, yet low amounts are present in cytosol and nucleus. Here we examined several aspects concerning the molecular function of yeast Nfs1p as a model protein. First, we demonstrated that purified Nfs1p facilitates the in vitro assembly of Fe/S proteins by using cysteine as its specific substrate. Thus, eukaryotic Nfs1 is a functional orthologue of the bacterial cysteine desulfurase IscS. Second, we showed that only the mitochondrial version but not the extramitochondrial version of Nfs1p is functional in generating cytosolic and nuclear Fe/S proteins. Mutation of the nuclear targeting signal of Nfs1p did not affect the maturation of cytosolic and nuclear Fe/S proteins, despite a severe growth defect under this condition. Nfs1p could not assemble an Fe/S cluster on the Isu scaffold proteins when they were located in the yeast cytosol. The lack of function of these central Fe/S cluster assembly components suggests that the maturation of extramitochondrial Fe/S protein does not involve functional copies of the mitochondrial Fe/S cluster assembly machinery in the yeast cytosol. Third, the extramitochondrial version of Nfs1p was shown to play a direct role in the thiomodification of tRNAs. Finally, we identified a highly conserved N-terminal β-sheet of Nfs1p as a functionally essential part of the protein. The implication of these findings for the structural stability of Nfs1p and for its targeting mechanism to mitochondria and cytosol/nucleus will be discussed.

Original languageEnglish
Pages (from-to)36906-36915
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number35
DOIs
Publication statusPublished - Aug 27 2004

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Protein S
Yeast
Saccharomyces cerevisiae
Cytosol
Mitochondria
Yeasts
Proteins
Transfer RNA
Sulfur
Scaffolds
Machinery
Cysteine
Iron
cysteine desulfurase
Defects
Mutation
Substrates
Growth

ASJC Scopus subject areas

  • Biochemistry

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Functional characterization of the eukaryotic cysteine desulfurase Nfs1p from Saccharomyces cerevisiae. / Mühlenhoff, Ulrich; Balk, Janneke; Richhardt, Nadine; Kaiser, Jens T.; Sipos, Katalin; Kispál, G.; Lill, Roland.

In: Journal of Biological Chemistry, Vol. 279, No. 35, 27.08.2004, p. 36906-36915.

Research output: Contribution to journalArticle

Mühlenhoff, Ulrich ; Balk, Janneke ; Richhardt, Nadine ; Kaiser, Jens T. ; Sipos, Katalin ; Kispál, G. ; Lill, Roland. / Functional characterization of the eukaryotic cysteine desulfurase Nfs1p from Saccharomyces cerevisiae. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 35. pp. 36906-36915.
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