DNA mismatch repair and Chk1-dependent centrosome amplification in response to DNA alkylation damage.
Centrosome amplification is frequently observed in tumor cells exposed to genotoxic stress, however the underlying mechanisms and biological consequences are poorly understood. Here, we show that the anti-metabolite and alkylating agent 6-thioguanine (6-TG) induces centrosome amplification resulting in the formation of multi-polar spindles when damaged cells subsequently enter mitosis. These aberrant, multi-polar mitoses are frequently resolved by asymmetric cell divisions causing unequal segregation of genetic material and cell death in one or both daughter products. We show that this phenomenon is associated with transient cell cycle delay in S- and G(2)-phase and is dependent on DNA mismatch repair (DNA MMR) proficiency and Chk1 protein kinase activity. Although Chk1-deficient cells do not exhibit cell cycle delay, centrosome amplification, or multi-polar spindle formation, continued cell cycle progression in the presence of 6-TG eventually results in increased levels of mitotic catastrophe, most probably due to mitosis with incompletely replicated DNA. Taken together, these results reveal novel mechanisms of cell killing by 6-TG and underscore the importance of interactions between cell cycle checkpoints and DNA MMR in determining the fate of cells bearing DNA damage.
Robinson
Helen M R
HM
Beatson Institute for Cancer Research, Glasgow, UK. h.robinson@beatson.gla.ac.uk
Black
Elizabeth J
EJ
Brown
Robert
R
Gillespie
David A F
DA
D000477:Alkylating Agents / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D018385:Centrosome / Q000187:drug effects* / Q000502:physiology; D000071877:Checkpoint Kinase 1; D002645:Chickens; D004249:DNA Damage; D053843:DNA Mismatch Repair; D006801:Humans; D008954:Models, Biological; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology*; D008941:Spindle Apparatus / Q000187:drug effects; D013866:Thioguanine / Q000633:toxicity; D014407:Tumor Cells, Cultured
D000477:Alkylating Agents; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D013866:Thioguanine