Inhibition of Chk1-dependent G2 DNA damage checkpoint radiosensitizes p53 mutant human cells.
Cell cycle checkpoints are surveillance mechanisms that monitor and coordinate the order and fidelity of cell cycle events. When defects in the division program of a cell are detected, checkpoints prevent the pursuant cell cycle transition through regulation of the relevant cyclin-cdk complex(es). Checkpoints that respond to DNA damage have been described for the G1, S and G2 phases of the cell cycle. The p53 tumour suppressor is a key regulator of G1/S checkpoints, and can promote cell cycle delay or apoptosis in response to DNA damage. The importance of these events to cellular physiology is highlighted by the fact that tumours, in which p53 is frequently mutated, have widespread defects in the G1/S DNA damage checkpoints and a heightened level of genomic instability. G2/M DNA damage checkpoints have been defined by yeast genetics, though the genes in this response are conserved in mammals. We show here using biochemical and physiological assays that p53 is dispensable for a DNA damage checkpoint activated in the G2 phase of the cell cycle. Moreover, upregulation of p53 through serine 20 phosphorylation, does not occur in G2. Conversely, we show that the Chk1 protein kinase is essential for the human G2 DNA damage checkpoint. Importantly, inhibition of Chk1 in p53 deficient cells greatly sensitizes them to radiation, validating the hypothesis of targeting Chk1 in rational drug design and development for anti-cancer therapies.
Koniaras
K
K
Trescowthick Research Laboratories, Peter MacCallum Cancer Institute, A'Beckett St, Melbourne, VIC 8006 Australia.
Cuddihy
A R
AR
Christopoulos
H
H
Hogg
A
A
O'Connell
M J
MJ
D015153:Blotting, Western; D002453:Cell Cycle / Q000528:radiation effects; D002467:Cell Nucleus / Q000378:metabolism; D000071877:Checkpoint Kinase 1; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D016195:G2 Phase; D005799:Genes, Dominant; D016158:Genes, p53 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011836:Radiation Tolerance; D013997:Time Factors
D004247:DNA; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1