title: Mammalian ribonucleotide reductase subunit p53R2 is required for mitochondrial DNA replication and DNA repair in quiescent cells.
abstract: In postmitotic mammalian cells, protein p53R2 substitutes for protein R2 as a subunit of ribonucleotide reductase. In human patients with mutations in RRM2B, the gene for p53R2, mitochondrial (mt) DNA synthesis is defective, and skeletal muscle presents severe mtDNA depletion. Skin fibroblasts isolated from a patient with a lethal homozygous missense mutation of p53R2 grow normally in culture with an unchanged complement of mtDNA. During active growth, the four dNTP pools do not differ in size from normal controls, whereas during quiescence, the dCTP and dGTP pools decrease to 50% of the control. We investigate the ability of these mutated fibroblasts to synthesize mtDNA and repair DNA after exposure to UV irradiation. Ethidium bromide depleted both mutant and normal cells of mtDNA. On withdrawal of the drug, mtDNA recovered equally well in cycling mutant and control cells, whereas during quiescence, the mutant fibroblasts remained deficient. Addition of deoxynucleosides to the medium increased intracellular dNTP pools and normalized mtDNA synthesis. Quiescent mutant fibroblasts were also deficient in the repair of UV-induced DNA damage, as indicated by delayed recovery of dsDNA analyzed by fluorometric analysis of DNA unwinding and the more extensive and prolonged phosphorylation of histone H2AX after irradiation. Supplementation by deoxynucleosides improved DNA repair. Our results show that in nontransformed cells only during quiescence, protein p53R2 is required for maintenance of mtDNA and for optimal DNA repair after UV damage.
authors: Department of Biology,  University of Padova, I-35131 Padua,  Italy.
authors: Giovanna
authors: G
authors: Pontarin
authors: Paola
authors: P
authors: Ferraro
authors: Leonardo
authors: L
authors: Bee
authors: Peter
authors: P
authors: Reichard
authors: Vera
authors: V
authors: Bianchi
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004272:DNA, Mitochondrial / Q000235:genetics*; D004996:Ethidium / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology / Q000528:radiation effects; D005453:Fluorescence; D005470:Fluorometry; D006657:Histones / Q000378:metabolism; D006801:Humans; D006918:Hydroxyurea / Q000378:metabolism / Q000494:pharmacology; D008322:Mammals / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects / Q000528:radiation effects; D021122:Protein Subunits / Q000378:metabolism; D012264:Ribonucleotide Reductases / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: D018797:Cell Cycle Proteins; D004272:DNA, Mitochondrial; C468783:H2AX protein, human; D006657:Histones; D009711:Nucleotides; D021122:Protein Subunits; C405947:RRM2B protein, human; D012264:Ribonucleotide Reductases; D004996:Ethidium; D006918:Hydroxyurea