Break-induced ATR and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

TY - JOUR

T1 - Break-induced ATR and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

AU - Moss, Jennifer

AU - Tinline-Purvis, Helen

AU - Walker, Carol A

AU - Folkes, Lisa K

AU - Stratford, Michael R

AU - Hayles, Jacqueline

AU - Hoe, Kwang-Lae

AU - Kim, Dong-Uk

AU - Park, Han-Oh

AU - Kearsey, Stephen E

AU - Fleck, Oliver

AU - Holmberg, Christian

AU - Nielsen, Olaf

AU - Humphrey, Timothy C

PY - 2010/12/1

Y1 - 2010/12/1

N2 - Nucleotide synthesis is a universal response to DNA damage, but how this response facilitates DNA repair and cell survival is unclear. Here we establish a role for DNA damage-induced nucleotide synthesis in homologous recombination (HR) repair in fission yeast. Using a genetic screen, we found the Ddb1-Cul4Cdt2 ubiquitin ligase complex and ribonucleotide reductase (RNR) to be required for HR repair of a DNA double-strand break (DSB). The Ddb1-Cul4Cdt2 ubiquitin ligase complex is required for degradation of Spd1, an inhibitor of RNR in fission yeast. Accordingly, deleting spd1 + suppressed the DNA damage sensitivity and the reduced HR efficiency associated with loss of ddb1+ or cdt2+. Furthermore, we demonstrate a role for nucleotide synthesis in postsynaptic gap filling of resected ssDNA ends during HR repair. Finally, we define a role for Rad3 (ATR) in nucleotide synthesis and HR through increasing Cdt2 nuclear levels in response to DNA damage. Our findings support a model in which break-induced Rad3 and Ddb1-Cul4Cdt2 ubiquitin ligase-dependent Spd1 degradation and RNR activation promotes postsynaptic ssDNA gap filling during HR repair.

AB - Nucleotide synthesis is a universal response to DNA damage, but how this response facilitates DNA repair and cell survival is unclear. Here we establish a role for DNA damage-induced nucleotide synthesis in homologous recombination (HR) repair in fission yeast. Using a genetic screen, we found the Ddb1-Cul4Cdt2 ubiquitin ligase complex and ribonucleotide reductase (RNR) to be required for HR repair of a DNA double-strand break (DSB). The Ddb1-Cul4Cdt2 ubiquitin ligase complex is required for degradation of Spd1, an inhibitor of RNR in fission yeast. Accordingly, deleting spd1 + suppressed the DNA damage sensitivity and the reduced HR efficiency associated with loss of ddb1+ or cdt2+. Furthermore, we demonstrate a role for nucleotide synthesis in postsynaptic gap filling of resected ssDNA ends during HR repair. Finally, we define a role for Rad3 (ATR) in nucleotide synthesis and HR through increasing Cdt2 nuclear levels in response to DNA damage. Our findings support a model in which break-induced Rad3 and Ddb1-Cul4Cdt2 ubiquitin ligase-dependent Spd1 degradation and RNR activation promotes postsynaptic ssDNA gap filling during HR repair.

KW - Adaptor Proteins, Signal Transducing

KW - Cell Cycle Proteins

KW - DNA Breaks, Double-Stranded

KW - DNA Repair

KW - DNA-Binding Proteins

KW - Gene Deletion

KW - Nucleotides

KW - Protein Kinases

KW - Recombination, Genetic

KW - Ribonucleotide Reductases

KW - Schizosaccharomyces

KW - Schizosaccharomyces pombe Proteins

U2 - 10.1101/gad.1970810

DO - 10.1101/gad.1970810

M3 - Journal article

C2 - 21123655

SN - 0890-9369

VL - 24

SP - 2705

EP - 2716

JO - Genes & Development

JF - Genes & Development

IS - 23

ER -