title: Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA.
abstract: 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 Escherichia coli in vitro, albeit at lower efficiencies than with ATP. This work adds to the toolbox of nucleoside analogs useful for RNA labeling.
authors.affiliation: University of California, Davis, Department of Chemistry, One Shields Ave, Davis 95616, CA, USA.
authors.forename: Yuxuan
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: University of California, Davis, Department of Chemistry, One Shields Ave, Davis 95616, CA, USA. Electronic address: pabeal@ucdavis.edu.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Beal
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000480:Alkynes / Q000737:chemistry*; D017123:Bacteriophage T7 / Q000201:enzymology; D057930:Click Chemistry; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D011061:Poly A / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000480:Alkynes; D011061:Poly A; D012313:RNA; D000255:Adenosine Triphosphate; D012321:DNA-Directed RNA Polymerases
keywords: ATP analogs; Cellular RNA labeling; Click chemistry; Polyadenylation; T7 transcription