title: Conserved electron donor complex Dre2-Tah18 is required for ribonucleotide reductase metallocofactor assembly and DNA synthesis.
abstract: Eukaryotic ribonucleotide reductases (RNRs) require a diferric-tyrosyl radical (Fe(III)2-Y•) cofactor to produce deoxynucleotides essential for DNA replication and repair. This metallocofactor is an important target of RNR-based therapeutics, although mechanisms of in vivo cofactor assembly, inactivation, and reactivation are poorly understood. Here, we demonstrate that the conserved Fe-S protein-diflavin reductase complex, Dre2-Tah18, plays a critical role in RNR cofactor biosynthesis. Depletion of Dre2 affects both RNR gene transcription and mRNA turnover through the activation of the DNA-damage checkpoint and the Aft1/Aft2-controlled iron regulon. Under conditions of comparable RNR protein levels, cells with diminishing Dre2 have significantly reduced ability to make deoxynucleotides. Furthermore, the kinetics and levels of in vivo reconstitution of the RNR cofactor are severely impaired in two conditional tah18 mutants. Together, these findings provide insight into RNR cofactor formation and reveal a shared mechanism underlying assembly of the Fe(III)2-Y• cofactor in RNR and the Fe-S clusters in cytosolic and nuclear proteins.
authors.affiliation: Departments of Chemistry and Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Haoran
authors.initials: H
authors.lastname: Li
authors.forename: Caiguo
authors.initials: C
authors.lastname: Zhang
authors.forename: Xiuxiang
authors.initials: X
authors.lastname: An
authors.forename: Lili
authors.initials: L
authors.lastname: Liu
authors.forename: JoAnne
authors.initials: J
authors.lastname: Stubbe
authors.forename: Mingxia
authors.initials: M
authors.lastname: Huang
mesh_terms: D003067:Coenzymes / Q000378:metabolism; D004249:DNA Damage; D004271:DNA, Fungal / Q000096:biosynthesis*; D004583:Electrons; D007501:Iron / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010088:Oxidoreductases / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism; D012262:Ribonucleoside Diphosphate Reductase / Q000378:metabolism*; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013696:Temperature
chemical_list: D003067:Coenzymes; D004271:DNA, Fungal; C531900:Dre2 protein, S cerevisiae; D007506:Iron-Sulfur Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D007501:Iron; D010088:Oxidoreductases; C552878:Tah18 protein, S cerevisiae; D012262:Ribonucleoside Diphosphate Reductase; C523202:Rnr4 protein, S cerevisiae
keywords: S phase; dNTP pool; genome stability; iron cofactor; iron regulon
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