title: Visualization of ribonucleotide reductase catalytic oxidation establishes thioredoxins as its major reductants in yeast.
abstract: Thioredoxins and/or glutaredoxins assist ribonucleotide reductase, and other such enzymes that require disulfide bond reduction during their catalytic cycle. In Saccharomyces cerevisiae, the presence of either pathway is essential but which of these pathways operates in ribonucleotide reductase reduction and how this function contributes to the pathways' essential nature have not been definitively established. We have identified two in vivo redox forms of the S. cerevisiae ribonucleotide reductase R1 subunit, which correspond to catalytically reduced or oxidized enzymes. Cells lacking thioredoxins, which exhibit an elongated S phase, accumulate R1 in its oxidized form and also contain significantly decreased deoxyribonucleotide levels during the S phase. Overexpressing R1 in these cells increases both the amount of the R1 reduced form and the concentrations of deoxyribonucleotides and accelerates DNA replication. These results establish thioredoxins as the major RNR reducing system in yeast and indicate that impaired RNR reduction accounts for the S phase defects of thioredoxin-deficient cells.
authors.affiliation: CEA, DSV, DBJC, Service de Biochimie et de Génétique Moléculaire, 91191 Gif-sur-Yvette Cedex, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Camier
authors.forename: Emilie
authors.initials: E
authors.lastname: Ma
authors.forename: Christophe
authors.initials: C
authors.lastname: Leroy
authors.forename: Alain
authors.initials: A
authors.lastname: Pruvost
authors.forename: Michel
authors.initials: M
authors.lastname: Toledano
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: Marsolier-Kergoat
mesh_terms: D002384:Catalysis; D010084:Oxidation-Reduction; D010957:Plasmids; D012264:Ribonucleotide Reductases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*
chemical_list: D013879:Thioredoxins; D012264:Ribonucleotide Reductases