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

Jennifer Moss; Helen Tinline-Purvis; Carol A Walker; Lisa K Folkes; Michael R Stratford; Jacqueline Hayles; Kwang-Lae Hoe; Dong-Uk Kim; Han-Oh Park; Stephen E Kearsey; Oliver Fleck; Christian Holmberg; Olaf Nielsen; Timothy C Humphrey

doi:10.1101/gad.1970810

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

Jennifer Moss, Helen Tinline-Purvis, Carol A Walker, Lisa K Folkes, Michael R Stratford, Jacqueline Hayles, Kwang-Lae Hoe, Dong-Uk Kim, Han-Oh Park, Stephen E Kearsey, Oliver Fleck, Christian Holmberg, Olaf Nielsen, Timothy C Humphrey

Functional genomics

43 Citationer (Scopus)

Abstract

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-Cul4^Cdt2 ubiquitin ligase complex and ribonucleotide reductase (RNR) to be required for HR repair of a DNA double-strand break (DSB). The Ddb1-Cul4^Cdt2 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-Cul4^Cdt2 ubiquitin ligase-dependent Spd1 degradation and RNR activation promotes postsynaptic ssDNA gap filling during HR repair.

Originalsprog	Engelsk
Tidsskrift	Genes & Development
Vol/bind	24
Udgave nummer	23
Sider (fra-til)	2705-16
Antal sider	12
ISSN	0890-9369
DOI	https://doi.org/10.1101/gad.1970810
Status	Udgivet - 1 dec. 2010

Adgang til dokumentet

10.1101/gad.1970810

Citationsformater

Moss, J., Tinline-Purvis, H., Walker, C. A., Folkes, L. K., Stratford, M. R., Hayles, J., Hoe, K-L., Kim, D-U., Park, H-O., Kearsey, S. E., Fleck, O., Holmberg, C., Nielsen, O., & Humphrey, T. C. (2010). Break-induced ATR and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast. Genes & Development, 24(23), 2705-16. https://doi.org/10.1101/gad.1970810

Moss, J, Tinline-Purvis, H, Walker, CA, Folkes, LK, Stratford, MR, Hayles, J, Hoe, K-L, Kim, D-U, Park, H-O, Kearsey, SE, Fleck, O, Holmberg, C , Nielsen, O & Humphrey, TC 2010, 'Break-induced ATR and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast', Genes & Development, bind 24, nr. 23, s. 2705-16. https://doi.org/10.1101/gad.1970810

@article{baeead4c956e480f846493519445182f,

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

abstract = "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.",

keywords = "Adaptor Proteins, Signal Transducing, Cell Cycle Proteins, DNA Breaks, Double-Stranded, DNA Repair, DNA-Binding Proteins, Gene Deletion, Nucleotides, Protein Kinases, Recombination, Genetic, Ribonucleotide Reductases, Schizosaccharomyces, Schizosaccharomyces pombe Proteins",

author = "Jennifer Moss and Helen Tinline-Purvis and Walker, {Carol A} and Folkes, {Lisa K} and Stratford, {Michael R} and Jacqueline Hayles and Kwang-Lae Hoe and Dong-Uk Kim and Han-Oh Park and Kearsey, {Stephen E} and Oliver Fleck and Christian Holmberg and Olaf Nielsen and Humphrey, {Timothy C}",

year = "2010",

month = dec,

day = "1",

doi = "10.1101/gad.1970810",

language = "English",

volume = "24",

pages = "2705--16",

journal = "Genes & Development",

issn = "0890-9369",

publisher = "Cold Spring Harbor Laboratory Press",

number = "23",

}

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 -

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

Abstract

Adgang til dokumentet

Fingeraftryk

Citationsformater