Friedreich's ataxia (FRDA) is a debilitating cardio- and neurodegenerative disease currently affecting 6,000 Americans. Recent progress in FRDA research has linked pathogenesis of FRDA to the absence of Frataxin (Fxn), a protein that regulates iron-sulfur cluster (Fe-S) biosynthesis, possibly by serving as a regulator of the pathway or as an iron chaperone that delivers metal to protein partners in the Fe-S cluster bioassembly (ISC) pathway. In eukaryotes, the predominant ISC assembly pathway is located within the mitochondria and driven by coordinated association of the bioassembly scaffold protein (Isu1), a cysteine desulfurase that provides atomic sulfur (Nfs1), an accessory protein (Isd11) and frataxin (Yfh1). The long-range goal of the Stemmler laboratory is to characterize the molecular details of the Fe-S cluster assembly pathway in eukaryotes. The objective of this proposal is to characterize the active site of the Isu1 scaffolding protein in addition to the overll ISC assembly complex. The findings of this study will help in the development strategies to augment the activity of this pathway that is deficient when frataxin is absent. All studies will be performed in the yeast system, where the genetics of Fe-S cluster assembly have been well established. Our preliminary hypothesis is that Fxn provides two Fe (II) atoms and Nfs provides two sulfur atoms at distinct sites on Isu1. Subsequent Isu1 rearrangement brings both sulfur and Fe (II) species together at Isu1's active site to accomplish cluster assembly. This proposal follows two specific aims. Firstly, Isu1 will be manipulated to identify and characterize critical residues essential for Fe-S assembly. Secondly, a multi-protein expression plasmid, with all 4 ISC complex proteins present, will be utilized to determine how Isu1, Nfs1, Isd11 assemble in the presence/absence of frataxin. As a whole, this proposal will facilitate the development of novel therapeutic strategies for FRDA.

Public Health Relevance

In recent years, several neurodegenerative disorders have been linked to dysfunctional iron metabolism with resulting iron accumulation and increased oxidative stress, both of which lead to the impaired health and eventually contribute to patient death. Relevant disorders include Alzheimer's Disease, Parkinson's Disease, Huntington's Disease and Friedreich's Ataxia, the most common form of hereditary ataxia. Friedreich's Ataxia is typically caused by a trinucleotide repeat expansion the first intron of the gene for the protei frataxin, leading to a reduction in protein production. Phenotypes of the disorder include general disruption in cellular iron homeostasis, mitochondrial iron overload and a breakdown in Fe-S cluster biosynthesis. This proposal will provide mechanistic details of the Fe-S cluster assembly process as it occurs in eukaryotes and help explain why it is dysfunctional in the absence of frataxin. This research plan is relevant to public health because it will facilitate the developmen of new therapeutic strategies for the treatment of Friedreich's Ataxia and will provide insight int deficient pathways in several other neurodegenerative disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
Project #
5F30DK101230-04
Application #
9392545
Study Section
Special Emphasis Panel (ZDK1)
Program Officer
Roy, Cindy
Project Start
2014-12-01
Project End
2018-11-30
Budget Start
2017-12-01
Budget End
2018-11-30
Support Year
4
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Wayne State University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Dzul, Stephen P; Rocha, Agostinho G; Rawat, Swati et al. (2017) In vitro characterization of a novel Isu homologue from Drosophila melanogaster for de novo FeS-cluster formation. Metallomics 9:48-60
Truong, Phan T; Gale, Eric M; Dzul, Stephen P et al. (2017) Steric Enforcement about One Thiolate Donor Leads to New Oxidation Chemistry in a NiSOD Model Complex. Inorg Chem 56:7761-7780
Steiner, Ramsey A; Dzul, Stephen P; Stemmler, Timothy L et al. (2017) Synthesis and Speciation-Dependent Properties of a Multimetallic Model Complex of NiSOD That Exhibits Unique Hydrogen-Bonding. Inorg Chem 56:2849-2862
Barupala, Dulmini P; Dzul, Stephen P; Riggs-Gelasco, Pamela Jo et al. (2016) Synthesis, delivery and regulation of eukaryotic heme and Fe-S cluster cofactors. Arch Biochem Biophys 592:60-75
Walter, Melody R; Dzul, Stephen P; Rodrigues, Andria V et al. (2016) Synthesis of Co(II)-NO(-) Complexes and Their Reactivity as a Source of Nitroxyl. J Am Chem Soc 138:12459-71
Plegaria, Jefferson S; Dzul, Stephen P; Zuiderweg, Erik R P et al. (2015) Apoprotein Structure and Metal Binding Characterization of a de Novo Designed Peptide, ?3DIV, that Sequesters Toxic Heavy Metals. Biochemistry 54:2858-73
Bafaro, Elizabeth M; Antala, Sagar; Nguyen, Tuong-Vi et al. (2015) The large intracellular loop of hZIP4 is an intrinsically disordered zinc binding domain. Metallomics 7:1319-30