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首页> 外文期刊>Biochemistry >Incorporation of CC steps into Z-DNA: Interplay between B-Z junction and Z-DNA helical formation
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Incorporation of CC steps into Z-DNA: Interplay between B-Z junction and Z-DNA helical formation

机译：将CC步骤并入Z-DNA：B-Z连接和Z-DNA螺旋形成之间的相互作用

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The left-handed DNA structure, Z-DNA, is believed to play important roles in gene expression and regulation. Z-DNA forms sequence-specifically with a preference for sequences rich in pyrimidine/purine dinucleotide steps. In vivo, Z-DNA is generated in the presence of negative supercoiling or upon binding proteins that absorb the high energetic cost of the B-to-Z transition, including the creation of distorted junctions between B-DNA and Z-DNA. To date, the sequence preferences for the B-to-Z transition have primarily been studied in the context of sequence repeats lacking B-Z junctions. Here, we develop a method for characterizing sequence-specific preferences for Z-DNA formation and B-Z junction localization within heterogeneous DNA duplexes that is based on combining 2-aminopurine fluorescence measurements with a new quantitative application of circular dichroism spectroscopy for determining the fraction of B- versus Z-DNA. Using this approach, we show that at least three consecutive CC dinucleotide steps, traditionally thought to disfavor Z-DNA, can be incorporated within heterogeneous ZDNA containing B-Z junctions upon binding to the Za domain of the RNA adenosine deaminase protein. Our results indicate that the incorporation of CC steps into Z-DNA is driven by favorable sequence-specific Z-Z and B-Z stacking interactions as well as by sequence-specific energetics that localize the distorted B-Z junction at flexible sites. Together, our results expose higher-order complexities in the Z-DNA code within heterogeneous sequences and suggest that Z-DNA can in principle propagate into a wider range of genomic sequence elements than previously thought.
机译：左手DNA结构Z-DNA被认为在基因表达和调控中起重要作用。 Z-DNA特异性地形成序列，优选富含嘧啶/嘌呤二核苷酸步骤的序列。在体内，Z-DNA是在负超螺旋的存在下产生的，或者是结合蛋白吸收B到Z过渡的高能量成本（包括在B-DNA和Z-DNA之间形成扭曲的连接）而生成的。迄今为止，主要是在缺乏B-Z连接的重复序列的背景下研究了B-Z过渡的序列偏好。在这里，我们开发了一种方法，用于表征异质DNA双链体内Z-DNA形成和BZ连接定位的序列特异性偏好，该方法基于将2-氨基嘌呤荧光测量与圆二色光谱的新定量应用相结合来确定B的分数的方法-与Z-DNA。使用这种方法，我们表明，传统上认为不赞成Z-DNA的至少三个连续的CC二核苷酸步骤可以在与RNA腺苷脱氨酶蛋白的Za结构域结合后并入含有B-Z连接的异质ZDNA中。我们的结果表明，CC步骤掺入Z-DNA的过程是由有利的序列特异性Z-Z和B-Z堆积相互作用以及序列特异性的能量学驱动的，这些能量将扭曲的B-Z连接定位在柔性位点。总之，我们的研究结果揭示了异质序列内Z-DNA代码的更高阶复杂性，并表明Z-DNA原则上可以传播到比以前认为的范围更广的基因组序列元素中。

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