DNA end resection, homologous recombination and DNA damage checkpoint activation require CDK1.
A single double-strand break (DSB) induced by HO endonuclease triggers both repair by homologous recombination and activation of the Mec1-dependent DNA damage checkpoint in budding yeast. Here we report that DNA damage checkpoint activation by a DSB requires the cyclin-dependent kinase CDK1 (Cdc28) in budding yeast. CDK1 is also required for DSB-induced homologous recombination at any cell cycle stage. Inhibition of homologous recombination by using an analogue-sensitive CDK1 protein results in a compensatory increase in non-homologous end joining. CDK1 is required for efficient 5' to 3' resection of DSB ends and for the recruitment of both the single-stranded DNA-binding complex, RPA, and the Rad51 recombination protein. In contrast, Mre11 protein, part of the MRX complex, accumulates at unresected DSB ends. CDK1 is not required when the DNA damage checkpoint is initiated by lesions that are processed by nucleotide excision repair. Maintenance of the DSB-induced checkpoint requires continuing CDK1 activity that ensures continuing end resection. CDK1 is also important for a later step in homologous recombination, after strand invasion and before the initiation of new DNA synthesis.
Ira
Grzegorz
G
Rosenstiel Center and Department of Biology, Brandeis University, Waltham, Massachusetts 02454-9110, USA.
Pellicioli
Achille
A
Balijja
Alitukiriza
A
Wang
Xuan
X
Fiorani
Simona
S
Carotenuto
Walter
W
Liberi
Giordano
G
Bressan
Debra
D
Wan
Lihong
L
Hollingsworth
Nancy M
NM
Haber
James E
JE
Foiani
Marco
M
D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000378:metabolism; D064447:Checkpoint Kinase 2; D004249:DNA Damage / Q000187:drug effects; D004260:DNA Repair / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal / Q000235:genetics; D000072235:Mating Factor; D015739:Nocodazole / Q000494:pharmacology; D010455:Peptides / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; D000072235:Mating Factor; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; D016203:CDC2 Protein Kinase; C067022:RAD53 protein, S cerevisiae; D015739:Nocodazole