Defective repair of mitomycin C crosslinks in Fanconi's anemia and loss in confluent normal human and xeroderma pigmentosum cells.
Crosslink repair of mitomycin C-induced interstrand crosslinks was studied in exponentially growing and confluent normal human, transformed W138CT-1, Fanconi's anemia (FA) and xeroderma pigmentosum (XP) group-A fibroblasts by the assay methods of alkaline sucrose centrifugation, hydroxyapatite column chromatography and S1-nuclease digestion. These three methods demonstrated unequivocally that crosslinking occurred at a rate of 0.13 crosslinks/10(8) Da per microgram per ml mitomycin C (less than or equal to 10 micrograms/ml) and the first half-excision of crosslinks followed the rapid first-order kinetics of 2-3 h half-life in exponentially-growing normal, WI38CT-1 and XP group-A cells. However, the first half-excision was completely defective in three out of the four FA strains tested and severely retarded in an FA strain. These results strongly support our previous observations in different strains of normal human, FA and XP group-A cells. An important new addition is that confluent, otherwise proficient, normal and XP cells almost completely lost the ability of the first, rapid half-excision of mitomycin C crosslinks in their DNA. This probably suggests that the enzyme or regulatory factor responsible for the half-excision, which differs from that for nucleotide excision repair, present constitutively in confluent cells, may be induced or activated only in the cycling cells. However, its relation to a defective FA factor is not clear at present.
D000741:Anemia, Aplastic / Q000503:physiopathology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002453:Cell Cycle; D004247:DNA / Q000302:isolation & purification; D004260:DNA Repair; D005199:Fanconi Anemia / Q000503:physiopathology*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D006801:Humans; D016685:Mitomycin; D008937:Mitomycins / Q000494:pharmacology*; D009691:Nucleic Acid Denaturation; D009695:Nucleic Acid Renaturation; D014983:Xeroderma Pigmentosum / Q000503:physiopathology*
D000903:Antibiotics, Antineoplastic; D008937:Mitomycins; D016685:Mitomycin; D004247:DNA