Pyrene is highly emissive when attached to the RNA duplex but not to the DNA duplex: the structural basis of this difference.
Pyrene is highly emissive when attached to the RNA duplex but not to the DNA duplex: the structural basis of this difference.
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DOI：
10.1093/nar/gki889
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发表时间：
2005
影响因子：
14.9
通讯作者：
Yamana K
中科院分区：
文献类型：
--
作者：
Nakamura M;Fukunaga Y;Sasa K;Ohtoshi Y;Kanaori K;Hayashi H;Nakano H;Yamana K
通过对通过糖残基处的一个碳连接体共价连接到芘基团的寡核苷酸进行结合和荧光研究，我们之前发现芘修饰的RNA寡核苷酸在单链形式下不能很好地发射，但是连接的芘在与互补RNA链结合时以显着高的量子产率发射。与此形成鲜明对比的是，类似修饰的芘-DNA 探针在双链和单链形式下都表现出非常弱的荧光。因此，芘修饰的RNA寡核苷酸为监测RNA杂交提供了有用的工具。本文的目的是介绍芘修饰的 RNA/RNA 和芘修饰的 DNA/DNA 双链体不同荧光特性的结构基础。吸收、荧光各向异性和圆二色性研究的结果一致表明，附着在RNA双链体上的芘位于双链体的外部，而掺入DNA双链体中的芘则插入双螺旋中。 1H NMR 测量明确证实连接到 DNA 双链体的芘确实插入到双链体的碱基对之间。分子动力学模拟支持芘-RNA/RNA 和芘-DNA/DNA 双链体之间芘周围局部结构元件的这些差异。
Through binding and fluorescence studies of oligonucleotides covalently attached to a pyrene group via one carbon linker at the sugar residue, we previously found that pyrene-modified RNA oligonucleotides do not emit well in the single-stranded form, yet the attached pyrene emits with a significantly high quantum yield upon binding to a complementary RNA strand. In sharp contrast, similarly modified pyrene–DNA probes exhibit very weak fluorescence both in the double-stranded and single-stranded forms. The pyrene-modified RNA oligonucleotides therefore provide a useful tool for monitoring RNA hybridization. The purpose of this paper is to present the structural basis for the different fluorescence properties of pyrene-modified RNA/RNA and pyrene-modified DNA/DNA duplexes. The results of absorption, fluorescence anisotropy and circular dichroism studies all consistently indicated that the pyrene attached to the RNA duplex is located outside of the duplex, whereas the pyrene incorporated into the DNA duplex intercalates into the double helix. 1H NMR measurements unambiguously confirmed that the pyrene attached to the DNA duplex indeed intercalates between the base pairs of the duplex. Molecular dynamics simulations support these differences in the local structural elements around the pyrene between the pyrene–RNA/RNA and the pyrene–DNA/DNA duplexes.
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