Inhibition of DNA polymerase reaction by pyrimidine nucleotide analogues lacking the 2-keto group

Guo, M. J.; Hildbrand, S.; Leumann, Christian; McLaughlin, L. W.; Waring, M. J. (1998). Inhibition of DNA polymerase reaction by pyrimidine nucleotide analogues lacking the 2-keto group. Nucleic acids research, 26(8), pp. 1863-1869. Oxford University Press 10.1093/nar/26.8.1863

	Text 1863.full.pdf - Published Version Restricted to registered users only Available under License Publisher holds Copyright. Download (122kB) | Request a copy
Preview	Text 26-8-1863.pdf - Other Available under License Publisher holds Copyright. Download (138kB) | Preview

To investigate the influence of the pyrimidine 2-keto group on selection of nucleotides for incorporation into DNA by polymerases, we have prepared two C nucleoside triphosphates that are analogues of dCTP and dTTP, namely 2-amino-5-(2'-deoxy-beta-d-ribofuranosyl)pyridine-5'-triphosphate (d*CTP) and 5-(2'-deoxy- beta-d-ribofuranosyl)-3-methyl-2-pyridone-5'-triphosphate (d*TTP) respectively. Both proved strongly inhibitory to PCR catalysed by Taq polymerase; d*TTP rather more so than d*CTP. In primer extension experiments conducted with either Taq polymerase or the Klenow fragment of Escherichia coli DNA polymerase I, both nucleotides failed to substitute for their natural pyrimidine counterparts. Neither derivative was incorporated as a chain terminator. Their capacity to inhibit DNA polymerase activity may well result from incompatibility with the correctly folded form of the polymerase enzyme needed to stabilize the transition state and catalyse phosphodiester bond formation.

Interest & Impact

Downloads

88 since deposited on 20 May 2015
27 in the past 12 months

Citations

5 Citations in Web of Science ®
6 Citations in Scopus

Search

in Google Scholar™

Services

Actions (login required)

Edit item

Actions (login required)

Edit item