Spontaneous and cisplatin-induced recombination in Escherichia coli
Graduate School of Biomedical Sciences; Department of Biochemistry and Molecular Pharmacology
Medical Subject Headings
Cell Survival; Cisplatin; Cross-Linking Reagents; DNA Repair; DNA, Bacterial; Escherichia coli; Escherichia coli Proteins; Gene Deletion; Gene Silencing; Genes, Bacterial; Lac Operon; Models, Genetic; Recombination, Genetic
Life Sciences | Medicine and Health Sciences
To measure cisplatin (cis-diaminodichloroplatinum(II))-induced recombination, we have used a qualitative intrachromosomal assay utilizing duplicate inactive lac operons containing non-overlapping deletions and selection for Lac+ recombinants. The two operons are separated by one Mb and conversion of one of them yields the Lac+ phenotype. Lac+ formation for both spontaneous and cisplatin-induced recombination requires the products of the recA, recBC, ruvA, ruvB, ruvC, priA and polA genes. Inactivation of the recF, recO, recR and recJ genes decreased cisplatin-induced, but not spontaneous, recombination. The dependence on PriA and RecBC suggests that recombination is induced following stalling or collapse of replication forks at DNA lesions to form double strand breaks. The lack of recombination induction by trans-DDP suggests that the recombinogenic lesions for cisplatin are purine-purine intrastrand crosslinks.
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Citation: DNA Repair (Amst). 2004 Jul 2;3(7):719-28. Link to article on publisher's site
Nowosielska, Anetta; Calmann, Melissa A.; Zdraveski, Zoran Z.; Essigmann, John M.; and Marinus, Martin G., "Spontaneous and cisplatin-induced recombination in Escherichia coli" (2004). GSBS Student Publications. 908.