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PMID: 12970194 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A novel eukaryotic factor for cytosolic Fe-S cluster assembly.

The EMBO journal ·Vol. 22 ·No. 18 ·2003-09-15 ·Pages 4826-35

Roy A, Solodovnikova N, Nicholson T, Antholine W, Walden WE

Abstract

Iron regulatory protein 1 (IRP1) is regulated through the assembly/disassembly of a [4Fe-4S] cluster, which interconverts IRP1 with cytosolic aconitase. A genetic screen to isolate Saccharomyces cerevisiae strains bearing mutations in genes required for the conversion of IRP1 to c-aconitase led to the identification of a previously uncharacterized, essential gene, which we call CFD1 (cytosolic Fe-S cluster deficient). CFD1 encodes a highly conserved, putative P-loop ATPase. A non-lethal mutation of CFD1 (cfd1-1) reduced c-aconitase specific activity in IRP1-transformed yeast by >90%, although IRP1 in these cells could be readily converted to c-aconitase in vitro upon incubation with iron alone. IRP1-transformed cfd1-1 yeast lacked EPR-detectable Fe-S clusters in c-aconitase, pointing to a defect in Fe-S cluster assembly. The specific activity of another cytosolic Fe-S protein, Leu1p, was also inhibited by >90% in cfd1-1 yeast, whereas activity of mitochondrial Fe-S proteins was not inhibited. Consistent with a cytosolic site of activity, Cfd1p was localized in the cytoplasm. To our knowledge, Cfd1p is the first cytoplasmic Fe-S cluster assembly factor described in eukaryotes.

MeSH Terms
Aconitate Hydratase/metabolism Amino Acid Sequence Animals Electron Spin Resonance Spectroscopy Gene Expression Regulation, Fungal Genetic Complementation Test Humans Iron Regulatory Protein 1/chemistry,genetics,metabolism Iron-Sulfur Proteins/chemistry,metabolism Molecular Sequence Data Recombinant Proteins/chemistry,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Sequence Alignment Sequence Homology, Amino Acid Transfection
Chemicals
Iron-Sulfur Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins Aconitate Hydratase Iron Regulatory Protein 1
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Roy Amit
Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL 60612, USA.
Solodovnikova Natalia
Nicholson Tracy
Antholine William
Walden William E
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg455
Published
2003-09-15
Pages
4826-35
Language
English
Region
England
NLM ID
8208664
PMCID
PMC212722
Subset
IM
Grants
NIDDK NIH HHS · R01 DK047281 · United States
NIDDK NIH HHS · DK47281 · United States
NCRR NIH HHS · RR01088 · United States
External Links
PubMed source DOI: 10.1093/emboj/cdg455 PMC Full Text View journal details
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