Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA

Yuxuan Zheng, Peter A. Beal

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1799-1802
Number of pages4
JournalBioorganic and Medicinal Chemistry Letters
Volume26
Issue number7
DOIs
StatePublished - 2016

Fingerprint

Alkynes
Nucleosides
Adenosine Triphosphate
RNA
Polynucleotide Adenylyltransferase
RNA Polymerase I
Azides
Cycloaddition
Cycloaddition Reaction
DNA-Directed RNA Polymerases
HeLa Cells
Labeling
Escherichia coli
Nucleic Acids
Copper
Substrates

Keywords

  • ATP analogs
  • Cellular RNA labeling
  • Click chemistry
  • Polyadenylation
  • T7 transcription

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Molecular Medicine
  • Organic Chemistry
  • Drug Discovery
  • Pharmaceutical Science

Cite this

Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA. / Zheng, Yuxuan; Beal, Peter A.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 26, No. 7, 2016, p. 1799-1802.

Research output: Contribution to journalArticle

@article{e3b96df48f8f43af9fd0f81a5261e5ee,
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.",
keywords = "ATP analogs, Cellular RNA labeling, Click chemistry, Polyadenylation, T7 transcription",
author = "Yuxuan Zheng and Beal, {Peter A.}",
year = "2016",
doi = "10.1016/j.bmcl.2016.02.038",
language = "English (US)",
volume = "26",
pages = "1799--1802",
journal = "Bioorganic and Medicinal Chemistry Letters",
issn = "0960-894X",
publisher = "Elsevier Limited",
number = "7",

}

TY - JOUR

T1 - Synthesis and evaluation of an alkyne-modified ATP analog for enzymatic incorporation into RNA

AU - Zheng, Yuxuan

AU - Beal, Peter A.

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - ATP analogs

KW - Cellular RNA labeling

KW - Click chemistry

KW - Polyadenylation

KW - T7 transcription

UR - http://www.scopus.com/inward/record.url?scp=84978959051&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84978959051&partnerID=8YFLogxK

U2 - 10.1016/j.bmcl.2016.02.038

DO - 10.1016/j.bmcl.2016.02.038

M3 - Article

C2 - 26927424

AN - SCOPUS:84978959051

VL - 26

SP - 1799

EP - 1802

JO - Bioorganic and Medicinal Chemistry Letters

JF - Bioorganic and Medicinal Chemistry Letters

SN - 0960-894X

IS - 7

ER -