DNA-DAMAGE BY ANTICANCER AGENTS RESOLVED AT THE NUCLEOTIDE LEVEL OF A SINGLE-COPY GENE - EVIDENCE FOR A NOVEL BINDING-SITE FOR CISPLATIN IN CELLS
DNA-DAMAGE BY ANTICANCER AGENTS RESOLVED AT THE NUCLEOTIDE LEVEL OF A SINGLE-COPY GENE - EVIDENCE FOR A NOVEL BINDING-SITE FOR CISPLATIN IN CELLS
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DOI：
10.1093/nar/22.12.2311
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发表时间：
1994-06-25
影响因子：
14.9
通讯作者：
HARTLEY, JA
中科院分区：
文献类型：
--
作者：
GRIMALDI, KA;MCADAM, SR;HARTLEY, JA
一种新的基于PCR的技术已经被开发用于研究DNA损伤剂在分离的基因组DNA或药物处理的细胞中的单拷贝基因中形成加合物的序列选择性。单链连接PCR（sslig-PCR）表明，顺铂和氮mu反应与鸟嘌呤的N-ras片段具有不同的序列特异性类似于以前在质粒DNA中观察到的。在顺铂处理的细胞中，sslig-PCR证明了在分离的DNA中发现的所有加合物，并且具有相同的序列选择性，显示出对GG和AG位点的偏好。然而，在细胞中，在转录和非转录链上的两个出现的序列5 '-TACT-3'处观察到顺铂的DNA结合的额外位点。该序列不是顺铂的公认靶点，代表细胞中形成的新型加合物。
A new PCR based technique has been developed to investigate the sequence selectivity of adduct formation by DNA damaging agents in a single copy gene in isolated genomic DNA or in drug treated cells. Single-strand ligation PCR (sslig-PCR) demonstrated that cisplatin and nitrogen mustards reacted with guanine in an N-ras fragment with varying sequence specificity similar to that observed previously in plasmid DNA. In cisplatin-treated cells sslig-PCR demonstrated all the adducts found in isolated DNA and with the same sequence selectivity showing a preference for GG and AG sites. However, in cells an additional site of DNA binding of cisplatin was observed at the two occurrences of the sequence 5'-TACT-3' on the transcribed and non-transcribed strands. This sequence is not a recognised target for cisplatin and represents a novel adduct formed in cells.