Nowosielska Anetta, Calmann Melissa A, Zdraveski Zoran, Essigmann John M, Marinus M G
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation Street, LRB823 Worcester, MA 01655, USA.
DNA Repair (Amst). 2004 Jul 2;3(7):719-28. doi: 10.1016/j.dnarep.2004.02.009.
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.
为了检测顺铂（顺 - 二氨基二氯铂（II））诱导的重组，我们使用了一种定性的染色体内分析方法，该方法利用了两个含有非重叠缺失的重复无活性乳糖操纵子，并对Lac⁺重组体进行筛选。这两个操纵子相隔1兆碱基对，其中一个操纵子的转化会产生Lac⁺表型。自发重组和顺铂诱导的重组产生Lac⁺都需要recA、recBC、ruvA、ruvB、ruvC、priA和polA基因的产物。recF、recO、recR和recJ基因的失活降低了顺铂诱导的重组，但不影响自发重组。对PriA和RecBC的依赖性表明，在DNA损伤处复制叉停滞或崩溃形成双链断裂后诱导了重组。反式 - 顺铂不能诱导重组，这表明顺铂的重组性损伤是嘌呤 - 嘌呤链内交联。
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