Nuclear Localization of Yeast Nfs1p Is Required for Cell Survival*

Saccharomyces cerevisiae Nfs1p is mainly found in the mitochondrial matrix and has been shown to participate in iron-sulfur cluster assembly. We show here that Nfs1p contains a potential nuclear localization signal, RRRPR, in its mature part. When this sequence was mutated to RRGSR, the mutant protein could not restore cell growth under chromosomal NFS1-depleted conditions. However, this mutation did not affect the function of Nfs1p in biogenesis of mitochondrial iron-sulfur proteins. The growth defect of the mutant was complemented by simultaneous expression of the mature Nfs1p, which contains the intact nuclear localization signal but lacks its mitochondrial-targeting presequence. These results suggest that a fraction of Nfs1p is localized in the nucleus and is essential for cell viability.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  P. Kiley,et al.  The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Downs,et al.  Metabolic Defects Caused by Mutations in the isc Gene Cluster in Salmonella enterica Serovar Typhimurium: Implications for Thiamine Synthesis , 2000, Journal of bacteriology.

[4]  C. T. Lauhon,et al.  The iscS Gene in Escherichia coli Is Required for the Biosynthesis of 4-Thiouridine, Thiamin, and NAD* , 2000, The Journal of Biological Chemistry.

[5]  T. Hoshino,et al.  Contribution of Cysteine Desulfurase (NifS Protein) to the Biotin Synthase Reaction of Escherichia coli , 2000, Journal of bacteriology.

[6]  B. Bui,et al.  Enzyme-mediated sulfide production for the reconstitution of [2Fe-2S] clusters into apo-biotin synthase of Escherichia coli. Sulfide transfer from cysteine to biotin. , 2000, European journal of biochemistry.

[7]  C. T. Lauhon,et al.  Evidence for the Transfer of Sulfane Sulfur from IscS to ThiI during the in Vitro Biosynthesis of 4-Thiouridine inEscherichia coli tRNA* , 2000, The Journal of Biological Chemistry.

[8]  C. T. Lauhon,et al.  IscS is a sulfurtransferase for the in vitro biosynthesis of 4-thiouridine in Escherichia coli tRNA. , 1999, Biochemistry.

[9]  D. Pain,et al.  Yeast Mitochondrial Protein, Nfs1p, Coordinately Regulates Iron-Sulfur Cluster Proteins, Cellular Iron Uptake, and Iron Distribution* , 1999, The Journal of Biological Chemistry.

[10]  R. Lill,et al.  The Essential Role of Mitochondria in the Biogenesis of Cellular Iron-Sulfur Proteins , 1999, Biological chemistry.

[11]  M. Choder,et al.  Starved Saccharomyces cerevisiae Cells Have the Capacity to Support Internal Initiation of Translation* , 1999, The Journal of Biological Chemistry.

[12]  R. Lill,et al.  The mitochondrial proteins Atm1p and Nfs1p are essential for biogenesis of cytosolic Fe/S proteins , 1999, The EMBO journal.

[13]  D. Goldfarb,et al.  A Nuclear Export Signal Prevents Saccharomyces cerevisiae Hsp70 Ssb1p from Stimulating Nuclear Localization Signal-directed Nuclear Transport* , 1999, The Journal of Biological Chemistry.

[14]  B. Soltys,et al.  Mitochondrial-matrix proteins at unexpected locations: are they exported? , 1999, Trends in biochemical sciences.

[15]  N. Ueki,et al.  Selection system for genes encoding nuclear-targeted proteins , 1998, Nature Biotechnology.

[16]  T. Rouault,et al.  Targeting of a human iron-sulfur cluster assembly enzyme, nifs, to different subcellular compartments is regulated through alternative AUG utilization. , 1998, Molecular cell.

[17]  S. Garland,et al.  Suppressors of Superoxide Dismutase (SOD1) Deficiency in Saccharomyces cerevisiae , 1998, The Journal of Biological Chemistry.

[18]  W. Neupert,et al.  Import into Mitochondria, Folding and Retrograde Movement of Fumarase in Yeast* , 1998, The Journal of Biological Chemistry.

[19]  H. Hayashi,et al.  cDNA cloning and characterization of mouse nifS‐like protein, m‐Nfs1: mitochondrial localization of eukaryotic NifS‐like proteins , 1998, FEBS letters.

[20]  R. Klausner,et al.  YIpDCE1 - an integrating plasmid for dual constitutive expression in yeast. , 1998, Gene.

[21]  D. Dean,et al.  Assembly of Iron-Sulfur Clusters , 1998, The Journal of Biological Chemistry.

[22]  M K Johnson,et al.  Iron-sulfur proteins: new roles for old clusters. , 1998, Current opinion in chemical biology.

[23]  H. Beinert,et al.  Iron-sulfur clusters: nature's modular, multipurpose structures. , 1997, Science.

[24]  E. Phizicky,et al.  A 2′-Phosphotransferase Implicated in tRNA Splicing Is Essential in Saccharomyces cerevisiae * , 1997, The Journal of Biological Chemistry.

[25]  S. Le,et al.  PDGF2/c-sis mRNA Leader Contains a Differentiation-linked Internal Ribosomal Entry Site (D-IRES)* , 1997, The Journal of Biological Chemistry.

[26]  D. Cavener,et al.  Ultrabithorax and Antennapedia 5' untranslated regions promote developmentally regulated internal translation initiation , 1997, Molecular and cellular biology.

[27]  A. Barton,et al.  Purification, characterization, gene cloning, and expression of Saccharomyces cerevisiae redoxyendonuclease, a homolog of Escherichia coli endonuclease III. , 1997, Biochemistry.

[28]  D. A. Court,et al.  Mitochondrial and Cytosolic Branched-chain Amino Acid Transaminases from Yeast, Homologs of the myc Oncogene-regulated Eca39 Protein* , 1996, The Journal of Biological Chemistry.

[29]  D. Flint,et al.  Escherichia coli contains a protein that is homologous in function and N-terminal sequence to the protein encoded by the nifS gene of Azotobacter vinelandii and that can participate in the synthesis of the Fe-S cluster of dihydroxy-acid dehydratase. , 1996, The Journal of biological chemistry.

[30]  R. Brent,et al.  Correlation of two-hybrid affinity data with in vitro measurements , 1995, Molecular and cellular biology.

[31]  N. Grishin,et al.  Modeling of the spatial structure of eukaryotic ornithine decarboxylases , 1995, Protein science : a publication of the Protein Society.

[32]  D. Dean,et al.  Catalytic formation of a nitrogenase iron-sulfur cluster. , 1994, The Journal of biological chemistry.

[33]  R. White,et al.  Mechanism for the desulfurization of L-cysteine catalyzed by the nifS gene product. , 1994, Biochemistry.

[34]  B. Trumpower,et al.  Acidic regions of cytochrome c1 are essential for ubiquinol-cytochrome c reductase activity in yeast cells lacking the acidic QCR6 protein. , 1993, Journal of biochemistry.

[35]  J. Bolin,et al.  Nitrogenase metalloclusters: structures, organization, and synthesis , 1993, Journal of bacteriology.

[36]  Robert H. White,et al.  Cysteine desulfurase activity indicates a role for NIFS in metallocluster biosynthesis. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[37]  D. Sun,et al.  Cloning, nucleotide sequence, and regulation of the Bacillus subtilis nadB gene and a nifS-like gene, both of which are essential for NAD biosynthesis , 1993, Journal of bacteriology.

[38]  D. Söll,et al.  SPL1-1, a Saccharomyces cerevisiae mutation affecting tRNA splicing , 1993, Journal of bacteriology.

[39]  M. Scott,et al.  Homeotic gene Antennapedia mRNA contains 5'-noncoding sequences that confer translational initiation by internal ribosome binding. , 1992, Genes & development.

[40]  E. Phizicky,et al.  An enzyme from Saccharomyces cerevisiae uses NAD+ to transfer the splice junction 2'-phosphate from ligated tRNA to an acceptor molecule. , 1991, The Journal of biological chemistry.

[41]  Rodney Rothstein,et al.  Elevated recombination rates in transcriptionally active DNA , 1989, Cell.

[42]  M. W. Clark,et al.  The subnuclear localization of tRNA ligase in yeast , 1987, The Journal of cell biology.

[43]  H. Beinert,et al.  The role of iron in the activation-inactivation of aconitase. , 1983, The Journal of biological chemistry.

[44]  B. Lemire,et al.  Flavinylation of succinate: ubiquinone oxidoreductase from Saccharomyces cerevisiae. , 1995, Methods in enzymology.

[45]  N. Martin,et al.  How single genes provide tRNA processing enzymes to mitochondria, nuclei and the cytosol. , 1994, Biochimie.

[46]  G. Fink,et al.  Laboratory course manual for methods in yeast genetics , 1986 .

[47]  J. Lowenstein,et al.  [6] Aconitase from pig heart: [EC 4.2.13 Citrate (isocitrate) hydro-lyase]☆ , 1969 .