A-tract and (+)-CC-1065-induced bending of DNA. Comparison of structural features using non-denaturing gel analysis, hydroxyl-radical footprinting, and high-field NMR.
A-tract and (+)-CC-1065-induced bending of DNA. Comparison of structural features using non-denaturing gel analysis, hydroxyl-radical footprinting, and high-field NMR.
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A-tract 和 ( )-CC-1065 诱导的 DNA 弯曲。
DOI：
10.1021/bi00068a003
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Hurley,LH
中科院分区：
文献类型：
--
作者：
Sun,D;Lin,CH;Hurley,LH
1993年2月9日收到的修订稿摘要：(+)-CC-1065是泽尔链霉菌生产的一种具有生物活性的DNA反应性抗肿瘤抗生素。在之前的一项研究中，我们已经报告了(+)-CC-1065产生的DNA弯曲与A-链的内在联系有相似之处[Lin，C.H.，Sun，D.，&Hurley，L.H.(1991)Chem。Toxicol资源。4、21-26]。在这篇文章中，我们使用非变性凝胶分析、羟基自由基足迹和高场核磁共振的组合，为两种类型的弯曲之间的区别和连接在两种类型的弯曲中的重要性提供了证据。对于A-链，我们证明了弯曲的轨迹位于A-链的中心，用(+)-CC-1065修饰3‘端腺嘌呤后，该轨迹向3’侧移动了不到1个碱基对，弯曲幅度显著增加。例如5‘-AGTTA*或5’-GATTA*(其中*表示药物结合部位)的Fordrug结合序列，发现弯曲轨迹在两个胸腺嘧啶之间，并且弯曲集中在2碱基对序列而不是5碱基对序列上，这与A区的情况相同。A束固有弯曲和(+)-CC-1065诱导弯曲之间的一个重要区别是温度的影响。然而，如前所述，A束弯曲的幅度随着温度的降低而增加，对于药物诱导的5‘-AGTTA*的弯曲，弯曲幅度随着温度的升高而增加。药物修饰的5‘-AGTTA*序列的羟基自由基足迹显示以TT序列为中心的切割减少，这可能与小槽宽度的减少有关。在一项平行的研究中，非自互补的12-聚体双链(5/-GGCGGAGTTA*GG-3‘)’(5‘-CCTAACTC-CGCC-3’)(图2B)和相应的(+)-CC-1065-修饰的双链加合物通过一维和二维‘H核磁共振和NOESY限制的分子力学和动力学计算进行了彻底的研究。在室温下，12-聚双链和(+)-CC-1065-12-聚双链加合物在水溶液中始终保持反碱基取向的整体B型DNA。12-聚体双链及其药物修饰的加合物的18C核苷酸都有一个平均的C3‘-endo糖折叠。12-二聚体在16A核苷酸有独特的内部运动，位于共价修饰的腺嘌呤互补配对的3‘侧，在18C-19T步骤有一个主要的扭结。在与(+)-CC-1065共价键合后，18C附近的不连续性被捕获并进一步夸大。此外，12-聚双链加合物在8T到9T的台阶上表现出小槽的压缩和两侧的加宽，但在共价修饰部位表现得尤其突然。在结构上，12-聚双链加合物与弯曲的DNA结构有许多相似之处，后者与A-链有内在的联系。药物对DNA的主要扭曲位于共价修饰链的9T和10A步。(+)-CC-1065(图1)是泽尔链霉菌产生的一种非常有效的抗肿瘤抗生素(Hanka等人，1978)。(+)-CC-1065通过反应腺嘌呤的N3与双链DNA反应，形成与小沟中5-bp1区域重叠的共价加合物(Swenson等人，1982；Hurley等人，1984；(+)-CC-1065由CPI亚基(图1中的A亚基)和
Revised Manuscript Received February 9, 1993 abstract:(+)-CC-1065 is a biologically potent DNA-reactive antitumor antibiotic produced by Streptomyces zelensis. In a previous study we have reportedthat (+)-CC-1065 produces bending of DNA that has similarities to that intrinsically associated with A-tracts [Lin, C. H., Sun, D., & Hurley, L. H.(1991) Chem. Res. Toxicol. 4, 21-26]. In this article we provide evidence using a combination of non-denaturing gel analysis, hydroxyl-radical footprinting, and high-field NMR for both distinctions between the two types of bends andthe importance of junctions in both types of bends. For A-tracts we demonstrate that the locus of bending is at the center of an A-tract and that upon modification of the 3'adenine with (+)-CC-1065 this locus is moved less than 1 base pair to the 3'side, and the bending magnitude is significantly increased. Fordrug bonding sequences such as 5'-AGTTA* or 5'-GATTA*(where* denotes the drug bonding site), the locus of bending is found to be betweenthe two thymines, and the bending is focused over a 2-base-pair sequence rather than a 5-base-pair sequence, as is the case for the A-tract. An important distinction between an A-tract intrinsic bend and a (+)-CC-1065-induced bend is the effect of temperature. While, as shown previously, the magnitude of A-tract bending increases with decrease in temperature, for drug-induced bending of 5'-AGTTA* the bending magnitude increases with increased temperature. Hydroxylradical footprintingof the drug-modified 5'-AGTTA* sequence shows a decrease in cleavage centered around the TT sequence, which is presumably associated with a decrease in minor groove width. In a parallel study, the non-self-complementary 12-mer duplex (5/-GGCGGAGTTA* GG-3')'(5'-CCTAACTC-CGCC-3')(Figure 2B) and the corresponding (+)-CC-1065-modified duplex adduct were examined thoroughly by one-and two-dimensional'H NMR and NOESY restrained molecular mechanics and dynamics calculations. Both the 12-mer duplex and the (+)-CC-1065-12-mer duplex adduct maintain an overall B-form DNA with the anti base orientation throughout in aqueous solution at room temperature. The 18C nucleotide of both the 12-mer duplex and its drug-modified adduct has an average C3'-endo sugar pucker. The 12-mer duplex exhibits a unique internal motion at the 16A nucleotide, which is located to the 3'side of the complementary partner of the covalentlymodified adenine, and a major kink at the 18C-19T step. Following covalent bonding with (+)-CC-1065, the discontinuity around18C is entrapped and further exaggerated. In addition, the 12-mer duplex adduct displays a compression of the minor groove at the 8T to 9T step and widening on both sides, but especially abruptly at the covalent modification site. Structurally, the 12-mer duplex adduct bears many similarities to a bent DNA structure, which is intrinsically associated with A-tracts. The major drug-induced distortion on DNA is localized at the 9T and 10A step of the covalently modified strand. A truncated junction model for the drug-entrapped/induced bending of DNA is proposed, and a comparison to intrinsic A-tract bending is made.(+)-CC-1065 (Figure 1) is an extremely potent antitumor antibiotic produced by Streptomyces zelensis (Hanka et al., 1978).(+)-CC-1065 reacts with double-stranded DNA through N3 of a reactive adenine, forming a covalent adduct that overlaps a 5-bp1 region in the minor groove (Swenson et al., 1982; Hurley et al., 1984; Scahill et al., 1990).(+)-CC-1065 consists of a CPI subunit (subunit A in Figure 1) and
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( )-CC-1065-DNA 加合物的结构：有序水分子的关键作用以及对磷酸盐催化参与共价反应的影响。
影响因子：
2.9
作者：
Lin,CH;Beale,JM;Hurley,LH
通讯作者：
Hurley,LH
影响因子：
2.9
作者：
HURLEY, LH;LEE, CS;ARISTOFF, PA
通讯作者：
ARISTOFF, PA
d(GGAAAAAAGG)n、d(CCAAAAAACC)n 和 (CCAAAAAAGG)n 的电泳行为以及对 DNA 弯曲模型的影响。
影响因子：
14.9
作者：
R. Abagyan;V. Mironov;B. Chernov;V. P. Chuprina;A. V. Ulyanov
通讯作者：
A. V. Ulyanov
( )-CC-1065 产生类似于腺嘌呤/胸腺嘧啶束的 DNA 弯曲。
影响因子：
4.1
作者：
Lin,CH;Sun,DY;Hurley,LH
通讯作者：
Hurley,LH
( )-CC-1065 对 T4 DNA 连接酶活性的抑制：证明 ( )-CC-1065 对 DNA 的硬化和卷绕作用的重要性。
DOI：
--
发表时间：
1992
期刊：
Anti-cancer drug design
影响因子：
--
作者：
Sun,D;Hurley,LH
通讯作者：
Hurley,LH