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N 2 ‐Substituted 2′‐Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ
Author(s) -
Gowda A. S. Prakasha,
Lee Marietta,
Spratt Thomas E.
Publication year - 2017
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201611607
Subject(s) - dna polymerase , chemistry , polymerase , deoxyguanosine , dna , microbiology and biotechnology , reactivity (psychology) , substrate (aquarium) , in vivo , processivity , stereochemistry , dna polymerase i , biochemistry , polymerase chain reaction , biology , reverse transcriptase , gene , genetics , medicine , ecology , alternative medicine , pathology
N 2 ‐Alkyl‐2′‐deoxyguanosine triphosphate (N 2 ‐alkyl‐dGTP) derivatives with methyl, butyl, benzyl, or 4‐ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N 2 ‐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 N 2 ‐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 N 2 ‐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.