Characterization of bleomycin-mediated cleavage of a hairpin DNA library.
Characterization of bleomycin-mediated cleavage of a hairpin DNA library.
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DOI：
10.1021/bi400779r
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发表时间：
2013-08-06
期刊：
影响因子：
2.9
通讯作者：
Hecht SM
中科院分区：
文献类型：
--
作者：
Segerman ZJ;Roy B;Hecht SM
对BLM A5的研究是利用先前分离的发夹dna文库进行的，这些dna被发现与无金属BLM紧密结合。Fe（II）•BLM对发夹dna 3‘和5’臂的裂解能力进行了表征。强结合的DNA被发现是Fe•BLM a5介导的发夹DNA切割的有效底物。令人惊讶的是，发现blm介导的最普遍的切割位点是5 ‘ -AT-3 ’二核苷酸序列。当使用任意选择的DNA底物检测时，这种二核苷酸序列和其他通常不能被BLM很好地切割的序列在检查10个发夹DNA文库时是明显的。发夹DNA文库暴露于Fe•BLM A5共产生132个DNA切割位点。发夹dna的3 ‘臂和5 ’臂上存在多个切割位点，表明其中一些可能是双链切割事件。因此，开发了一种测定方法，用于测试发夹DNA遭受双链DNA损伤的倾向。用这种方法对一个发夹DNA进行了表征，结果与先前报道的双链DNA切割一致，但序列选择性与先前报道的不同。
A study of BLM A5 was conducted using a previously isolated library of hairpin DNAs found to bind strongly to metal free BLM. The ability of Fe(II)•BLM to effect cleavage on both the 3' and 5'-arms of the hairpin DNAs was characterized. The strongly bound DNAs were found to be efficient substrates for Fe•BLM A5-mediated hairpin DNA cleavage. Surprisingly, the most prevalent site of BLM-mediated cleavage was found to be the 5′-AT-3′ dinucleotide sequence. This dinucleotide sequence, and other sequences generally not cleaved well by BLM when examined using arbitrarily chosen DNA substrates, were apparent when examining the library of ten hairpin DNAs. In total, 132 sites of DNA cleavage were produced by exposure of the hairpin DNA library to Fe•BLM A5. The existence of multiple sites of cleavage on both the 3′- and 5′-arms of the hairpin DNAs suggested that some of these might be double-strand cleavage events. Accordingly, an assay was developed with which to test the propensity of the hairpin DNAs to undergo double-strand DNA damage. One hairpin DNA was characterized using this method, and gave results consistent with earlier reports of double-strand DNA cleavage, but with a sequence selectivity different from those reported previously.
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影响因子：
2.9
作者：
ABSALON, MJ;WU, W;STUBBE, J
通讯作者：
STUBBE, J
影响因子：
15
作者：
BOGER, DL;HONDA, T;COLLETTI, SL
通讯作者：
COLLETTI, SL
影响因子：
15
作者：
Leitheiser, CJ;Smith, KL;Hecht, SM
通讯作者：
Hecht, SM
影响因子：
15
作者：
Akiyama, Yoshitsugu;Ma, Qian;Hecht, Sidney M.
通讯作者：
Hecht, Sidney M.
影响因子：
56.9
作者：
KOZARICH, JW;WORTH, L;STUBBE, J
通讯作者：
STUBBE, J