Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex.
Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex.
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CC-1065 和特定寡脱氧核苷酸双链体之间加合物的表征。
DOI：
10.1093/nar/12.15.6159
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发表时间：
1984
影响因子：
14.9
通讯作者：
Wierenga,W
中科院分区：
文献类型：
--
作者：
Needham-VanDevanter,DR;Hurley,LH;Reynolds,VL;Theriault,NY;Krueger,WC;Wierenga,W
CC-1065是链霉菌产生的一种有效的抗肿瘤抗生素。该药物通过腺嘌呤的N-3共价结合，位于DNA的小凹槽内。先前的研究表明，CC-1065与DNA中的腺嘌呤反应产生热不稳定的产物，可用于揭示其序列特异性。这些研究还提供了对DNA序列（5 ' - cggagttaggggg -3 '）的深入了解，该序列应该以明确的方式结合CC-1065的一个分子。该序列在序列5 ‘ -TTA-3 ’中含有CC-1065腺嘌呤结合位点，与互补链一起化学合成。CC-1065与寡双链反应得到的加合物保持了B-DNA的形式，并具有与小牛胸腺DNA形成的CC-1065复合物相似的最终CD谱。上面的l4mer在5 '端用32p标记，与其互补链退火，与CC-1065反应并加热。在测序凝胶上评估药物介导的链断裂。在标记的链上发生一次断裂，形成一个以8.5mer迁移的片段；随后的哌啶处理产生了一个以7mer形式迁移的片段，这是已知的5 ‘ -TTA-3 ’序列中CC-1065在腺嘌呤上的结合所期望的大小。
CC-1065 is a potent antitumor antibiotic produced byStreptomyceszelensis. The drug binds covalently through N-3 of adenine and lies within the minor groove of DNA. Previous studies indicated that CC-1065 reacted with adenine in DNA to yield a thermally labile product that could be used to reveal its sequence specificity. These studies also provided insight into a DNA sequence (5′-CGGAGTTAGGGGCG-3′) which should bind one molecule of CC-1065 in an unambiguous manner. This sequence, which contains the CC-1065 adenine binding site within the sequence 5′-TTA-3′ was chemically synthesized together with the complementary strand. CC-1065 reacted with the oligoduplex to give an adduct that maintained the B-DNA form and had a final CD spectrum similar to those of the CC-1065 comp1exes formed with calf thymus DNA. The above l4mer was 5′ end-labelled with32P, annealed with its complementary strand, reacted with CC-1065 and heated. Drug-mediated strand breakage was evaluated on a sequencing gel. A single break occurred in the labelled strands to give a fragment that migrated as an 8.5mer; subsequent piperidine treatment produced a fragment that migrated as a 7mer, which is the size expected from the known binding of CC-1065 at adenine in 5′-TTA-3′ sequences.