Sequence-specific alkylation by Y-shaped and tandem hairpin pyrrole-imidazole polyamides
Sequence-specific alkylation by Y-shaped and tandem hairpin pyrrole-imidazole polyamides
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DOI：
10.1002/chem.200700571
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发表时间：
2008-01-01
影响因子：
4.3
通讯作者：
Sugiyama, Hiroshi
中科院分区：
文献类型：
--
作者：
Sasaki, Shunta;Bando, Toshikazu;Sugiyama, Hiroshi
为了延长发夹吡咯-咪唑（Py-Im）聚酰胺1的目标DNA序列长度，我们设计并合成了Y形串联发夹Py-Im聚酰胺2和3，它们具有1-（氯甲基）-5-羟基-1,2-二氢-3H-苯并[e]吲哚（seco-CBI）作为DNA烷基化部分。使用 5'-Texas-Red 标记的 465 碱基对 (bp) DNA 片段进行高分辨率变性聚丙烯酰胺凝胶电泳显示，缀合物 2 和 3 在纳摩尔浓度下烷基化了目标 DNA 序列的腺嘌呤。在两个 8 bp 匹配序列 5'-AATAACCA-3'（位点 A）和 5'-AAAATTCCA-3'（位点 C）的 3' 端缀合 2 烷基化腺嘌呤 N3，而缀合物 3 识别 465 bp DNA 片段中的一个 10bp 匹配序列 5'-AGAATAACCA-3'（位点 A）。这些结果表明 Y 形和串联发夹聚酰胺的 seco-CBI 缀合物已扩展其目标烷基化序列。
To extend the target DNA sequence length of the hairpin pyrrole-imidazole (Py-Im) polyamide 1, we designed and synthesized Y-shaped and tandem hairpin Py-Im polyamides 2 and 3, which possess 1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole (seco-CBI) as DNA-alkylating moieties. High-resolution denaturing polyacrylamide gel electrophoresis by using 5'-Texas-Red-labeled 465 base pair (bp) DNA fragments revealed that conjugates 2 and 3 alkylated the adenine of the target DNA sequences at nanomolar concentrations. Conjugate 2 alkylated adenine N3 at the 3' end of two 8 bp match sequences, 5'-AATAACCA-3' (site A) and 5'-AAATTCCA-3' (site C), while conjugate 3 recognized one 10bp match sequence, 5'-AGAATAACCA-3' (siteA) in the 465 bp DNA fragments. These results demonstrate that seco-CBI conjugates of Y-shaped and tandem hairpin polyamides have extended their target alkylation sequences.