Kowalska Joanna, Lewdorowicz Magdalena, Zuberek Joanna, Bojarska Elzbieta, Wojcik Jacek, Cohen Lean S, Davis Richard E, Stepinski Janusz, Stolarski Ryszard, Darzynkiewicz Edward, Jemielity Jacek
Department of Biophysics, Institute of Experimental Physics, Warsaw University, Warsaw, Poland.
Nucleosides Nucleotides Nucleic Acids. 2005;24(5-7):595-600. doi: 10.1081/ncn-200061915.
Nucleosides and oligonucleotides with an oxygen replaced by sulfur atom are an interesting class of compounds because of their improved stability toward enzymatic cleavage by nucleases. We have synthesized several dinucleotide mRNA cap analogs containing a phosphorothioate moiety in the alpha, beta, or gamma position of 5',5'-triphosphate chain [m7Gp(s)ppG, m7Gpp(s)pG, and m7Gppp(s)G]. These are the first examples of the biologically important 5'mRNA cap analogs containing a phosphorothioate moiety, and these compounds may be useful in a variety of biochemical and biotechnological applications. Incorporation of a sulfur atom in the alpha or gamma position within the dinucleotide cap analog was achieved using PSCl3 in a nucleoside phosphorylation reaction followed by coupling the phosphorothioate of nucleoside with a second nucleotide. Synthesis of cap analogs with the phosphorothioate moiety in beta position was performed using an organic phosphorothioate salt in a coupling reaction with an activated nucleotide. The structures of newly synthesized compounds was confirmed using MS and 1H and 31P NMR spectroscopy. We present here the results of preliminary studies on their interaction with translation initiation factor eIF4E and enzymatic hydrolysis with human and nematode DcpS scavengers.
氧原子被硫原子取代的核苷和寡核苷酸是一类有趣的化合物，因为它们对核酸酶的酶促切割具有更高的稳定性。我们合成了几种在5',5'-三磷酸链的α、β或γ位置含有硫代磷酸酯部分的二核苷酸mRNA帽类似物[m7Gp(s)ppG、m7Gpp(s)pG和m7Gppp(s)G]。这些是含有硫代磷酸酯部分的具有生物学重要性的5'mRNA帽类似物的首个实例，并且这些化合物可能在多种生化和生物技术应用中有用。在核苷磷酸化反应中使用PSCl3，然后将核苷的硫代磷酸酯与第二个核苷酸偶联，从而在二核苷酸帽类似物的α或γ位置引入硫原子。使用有机硫代磷酸酯盐与活化的核苷酸进行偶联反应，合成β位置含有硫代磷酸酯部分的帽类似物。使用质谱、1H和31P核磁共振光谱确认新合成化合物的结构。我们在此展示了它们与翻译起始因子eIF4E相互作用以及被人和线虫DcpS清除剂进行酶促水解的初步研究结果。
