Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA.
Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA.
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DOI：
10.1016/j.bmcl.2016.02.038
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发表时间：
2016-04-01
影响因子：
2.7
通讯作者：
Beal PA
中科院分区：
文献类型：
--
作者：
Zheng Y;Beal PA
炔修饰的核苷类似物由于能够参与铜催化的叠氮化/炔环加成(CuAAC)反应，在核酸定位以及功能和结构研究中是有用的。本文介绍了7-乙炔基-8-氮杂-7-去氮腺苷(7-EAATP)的合成及其酶促掺入RNA的方法。7-EAA的游离核苷被HeLa细胞摄取，并通过内源性RNA聚合酶结合到细胞RNA中。此外，在体外，7-EAATP是来自大肠杆菌的T7 RNA聚合酶和聚(A)聚合酶的底物，尽管效率低于ATP。这项工作增加了对RNA标记有用的核苷类似物的工具箱。
Alkyne-modified nucleoside analogs are useful for nucleic acid localization as well as functional and structural studies because of their ability to participate in copper-catalyzed azide/alkyne cycloaddition (CuAAC) reactions. Here we describe the synthesis of the triphosphate of 7-ethynyl-8-aza-7-deazaadenosine (7-EAATP) and the enzymatic incorporation of 7-EAA into RNA. The free nucleoside of 7-EAA is taken up by HeLa cells and incorporated into cellular RNA by endogenous RNA polymerases. In addition, 7-EAATP is a substrate for both T7 RNA polymerase and poly (A) polymerase from E. coli in vitro, albeit at lower efficiencies than with ATP. This work adds to the toolbox of nucleoside analogs useful for RNA labeling.