Hypersensitivity of Cockayne's syndrome cells to camptothecin is associated with the generation of abnormally high levels of double strand breaks in nascent DNA.
We report that fibroblasts from individuals with Cockayne's Syndrome (CS), an autosomal recessive disease exhibiting hypersensitivity to UV, are also hypersensitive to the killing action of camptothecin (CPT). In normal and CS cell lines the level of the protein-linked single strand DNA breaks (SSBs) induced by equal doses of CPT is similar, and these DNA breaks disappear within minutes of the removal of CPT. Thus, the toxicity of CPT does not correlate with the primary DNA lesions induced by the drug, and the hypersensitivity of CS cells cannot be explained by excessive topoisomerase I activity or by a defect in the enzyme ligation step. We have reported that CPT toxicity in normal cells is closely associated with the generation of double-strand DNA breaks (DSBs), predominantly at sites of DNA replication. The hypersensitivity of CS cells to CPT correlates closely with the much higher level of DSBs in nascent DNA than in normal cells. These DSBs are long-lived in all cells, but in CS many more (about 10-fold) remain 24 h after CPT removal and are presumably responsible for the higher frequency of chromosome aberrations in these cells. In CS as in normal cells aphidicolin prevents the generation of replication-related DSBs, suggesting that the movement of the DNA polymerase is necessary for the induction by CPT of the cytotoxic DSBs. Resistance to CPT and UV is restored to wild type in proliferating hybrids constructed between CS lines from two different complementation groups as is the abundance of replication-related DSBs. On the basis of this complementation we conclude that the UV and CPT sensitivities are distinct phenotypic traits arising from mutations in the CS A and B genes.
Squires
S
S
Mammalian Cell DNA Repair Research Group, Department of Zoology, Cambridge, United Kingdom.
Ryan
A J
AJ
Strutt
H L
HL
Johnson
R T
RT
D016590:Aphidicolin / Q000494:pharmacology; D002166:Camptothecin / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D003057:Cockayne Syndrome / Q000235:genetics*; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects*; D004249:DNA Damage; D004260:DNA Repair; D006801:Humans; D066298:In Vitro Techniques; D012313:RNA / Q000096:biosynthesis; D013997:Time Factors; D059004:Topoisomerase I Inhibitors; D014466:Ultraviolet Rays
D059004:Topoisomerase I Inhibitors; D016590:Aphidicolin; D012313:RNA; D004247:DNA; D002166:Camptothecin