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Zusammenfassung:
Upon exposure to the carcinogens N‐acetoxy‐N‐2‐acetylaminofluorene and 7‐bromomethylbenz[a]anthracene, which bind covalently to DNA, ether‐permeabilized (nucleotide‐permeable) Escherichia coli wild‐type cells responded with DNA excision repair. This repair was missing in mutants carrying defects in genes uvrA, uvrB and uvrC, whereas it was present in uvrD and several rec mutants.
Enzymic activities involved were identified by measuring repair polymerization and size reduction of denatured DNA.
1
An easily measurable effect in E. coli wild‐type cells was carcinogen‐induced repair polymerization. When initiated by N‐acetoxy‐N‐2‐acetylaminofluorene or 7‐bromomethyl‐benz[a]anthracene, it depended upon an ATP‐requiring step; CTP, GTP or UTP did not substitute for ATP. DNA repair synthesis was inhibited by p‐chloromercuribenzoate and quinacrine. In uvrA, uvrB and uvrC mutants no carcinogen‐stimulated DNA synthesis could be detected, indicating that steps involved in pyrimidine dimer excision are also involved in chemorepair. In recA, recB and recC mutant cells, repair synthesis was stimulated by the carcinogens to a normal extent. This evidence excludes the ATP‐dependent recB,C deoxyribonuclease and recA gene products as playing an important role in carcinogen‐induced excision repair. polA1 cells showed drastically reduced levels of repair polymerization, indicating that DNA polymerase I is the main polymerizing enzyme.
2
As determined by DNA size reduction in alkaline sucrose gradients, the arylalkylating carcinogens caused endonucleolytic cleavage of endogenous DNA in wild‐type cells. This incision step was most effectively performed in the presence of ATP; UTP, CTP and GTP were only slightly effective. Incision was inhibited by p‐chloromercuribenzoate and quinacrine. When exposed to the arylalkylating carcinogens, uvrA, uvrB and uvrC mutant cells did not perform the incision step in the presence of ATP, suggesting the involvement of the respective gene products in the initiation of chemorepair
