N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ

A. S.Prakasha Gowda, Marietta Lee, Thomas E. Spratt

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

N2-Alkyl-2′-deoxyguanosine triphosphate (N2-alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N2-Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-ethynylbenzyl-dG with wild-type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol κ were able to incorporate N2-4-ethynylbenzyl-dG into the nucleus. This is the first instance of a Y-family-polymerase-specific dNTP, and this method could be used to probe the activity of pol κ in vivo.

Original languageEnglish (US)
Pages (from-to)2628-2631
Number of pages4
JournalAngewandte Chemie - International Edition
Volume56
Issue number10
DOIs
StatePublished - Jan 1 2017

Fingerprint

DNA-Directed DNA Polymerase
DNA
Derivatives
Substrates
Azides
Fibroblasts
deoxyguanosine triphosphate

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)

Cite this

@article{2d3d6559f3b84ee184326bdfb8c848c9,
title = "N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ",
abstract = "N2-Alkyl-2′-deoxyguanosine triphosphate (N2-alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N2-Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-ethynylbenzyl-dG with wild-type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol κ were able to incorporate N2-4-ethynylbenzyl-dG into the nucleus. This is the first instance of a Y-family-polymerase-specific dNTP, and this method could be used to probe the activity of pol κ in vivo.",
author = "Gowda, {A. S.Prakasha} and Marietta Lee and Spratt, {Thomas E.}",
year = "2017",
month = "1",
day = "1",
doi = "10.1002/anie.201611607",
language = "English (US)",
volume = "56",
pages = "2628--2631",
journal = "Angewandte Chemie - International Edition",
issn = "1433-7851",
publisher = "John Wiley and Sons Ltd",
number = "10",

}

N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ. / Gowda, A. S.Prakasha; Lee, Marietta; Spratt, Thomas E.

In: Angewandte Chemie - International Edition, Vol. 56, No. 10, 01.01.2017, p. 2628-2631.

Research output: Contribution to journalArticle

TY - JOUR

T1 - N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ

AU - Gowda, A. S.Prakasha

AU - Lee, Marietta

AU - Spratt, Thomas E.

PY - 2017/1/1

Y1 - 2017/1/1

N2 - N2-Alkyl-2′-deoxyguanosine triphosphate (N2-alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N2-Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-ethynylbenzyl-dG with wild-type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol κ were able to incorporate N2-4-ethynylbenzyl-dG into the nucleus. This is the first instance of a Y-family-polymerase-specific dNTP, and this method could be used to probe the activity of pol κ in vivo.

AB - N2-Alkyl-2′-deoxyguanosine triphosphate (N2-alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N2-Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-ethynylbenzyl-dG with wild-type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol κ were able to incorporate N2-4-ethynylbenzyl-dG into the nucleus. This is the first instance of a Y-family-polymerase-specific dNTP, and this method could be used to probe the activity of pol κ in vivo.

UR - http://www.scopus.com/inward/record.url?scp=85011312045&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85011312045&partnerID=8YFLogxK

U2 - 10.1002/anie.201611607

DO - 10.1002/anie.201611607

M3 - Article

C2 - 28140505

AN - SCOPUS:85011312045

VL - 56

SP - 2628

EP - 2631

JO - Angewandte Chemie - International Edition

JF - Angewandte Chemie - International Edition

SN - 1433-7851

IS - 10

ER -