Johannsen Silke, Paulus Susann, Düpre Nicole, Müller Jens, Sigel Roland K O
Institute of Inorganic Chemistry, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
J Inorg Biochem. 2008 May-Jun;102(5-6):1141-51. doi: 10.1016/j.jinorgbio.2007.12.023. Epub 2008 Jan 8.
In vitro transcription by T7 RNA polymerase can be used to construct scaffolds for the one-dimensional arrangement of mercury(II) ions. In these constructs, the metal ions are located inside of RNA double helices. By replacing the amide protons of two oppositely located uracil residues of complementary strands, mercury(II) becomes coordinated in a linear fashion to form metal-ion mediated base pairs, analogous to the well-known thymine-Hg-thymine base pair in DNA. This is shown here by a combination of various experimental techniques, including NMR spectroscopy, dynamic light scattering, as well as UV and CD spectroscopy. A total of five different double helices, including both palindromic and non-palindromic RNA sequences and between two and twenty consecutive uracil residues, have been synthesized and shown to be able to incorporate mercury(II). The synthesis of r(GGAGU 20CUCC) demonstrates that T7 polymerase is capable of handling long continuous stretches of identical nucleotides, albeit at the cost of an increasing number of abortion products and longer oligonucleotide strands that need to be separated by polyacrylamide gel electrophoresis. This work introduces RNA into the group of nucleic acids that can form metal ion mediated base pairs. The use of such metal-modified nucleic acids has been envisaged in various fields of research, including the generation of molecular wires.
T7 RNA 聚合酶介导的体外转录可用于构建汞(II)离子一维排列的支架。在这些构建体中,金属离子位于 RNA 双螺旋内部。通过替换互补链中两个相对位置的尿嘧啶残基的酰胺质子,汞(II)以线性方式配位形成金属离子介导的碱基对,类似于 DNA 中著名的胸腺嘧啶 - 汞 - 胸腺嘧啶碱基对。这通过多种实验技术的组合得以证明,包括核磁共振光谱、动态光散射以及紫外和圆二色光谱。总共合成了五种不同的双螺旋,包括回文和非回文 RNA 序列以及含有两到二十个连续尿嘧啶残基的序列,并证明它们能够结合汞(II)。r(GGAGU20CUCC)的合成表明,T7 聚合酶能够处理长的连续相同核苷酸序列,尽管代价是流产产物数量增加以及需要通过聚丙烯酰胺凝胶电泳分离更长的寡核苷酸链。这项工作将 RNA 引入了能够形成金属离子介导碱基对的核酸类别。这种金属修饰的核酸已被设想用于包括分子导线生成在内的各种研究领域。