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Publication Detail

Title: The effect of divalent nickel (Ni2+) on in vitro DNA replication by DNA polymerase alpha.

Authors: Chin, Y E; Snow, E T; Cohen, M D; Christie, N T

Published In Cancer Res, (1994 May 1)

Abstract: The effects of the carcinogenic metal nickel on DNA polymerase alpha (pol alpha) activity and fidelity have been analyzed. In the absence of Mg2+, the presence of Ni2+ ions at concentrations below 0.25 mM gave rise to a dose-dependent activation of pol alpha as monitored by [3H]dTMP incorporation into an activated DNA template. The apparent Km for Ni(2+)-dependent pol alpha incorporation of dTTP was estimated to be 25 microM, which was about 10 times higher than the Km for Mg2+ (2.3 microM). Above 0.25 mM, Ni2+ caused a dose-dependent inhibition of pol alpha activity and the Ki was calculated to be 1.5 mM. Scatchard analyses showed that Ni2+ binds to affinity-purified pol alpha and associated proteins at two tight binding sites with a Kd of approximately 50 microM and at eight weak binding sites with a Kd of approximately 4 mM. In the presence of 2 mM Mg2+, the addition of Ni2+ to the reactions caused an inhibition of polymerase activity. The inhibition patterns tended to switch from competitive to mixed-type to noncompetitive as a function of Ni2+ concentration. Lastly, Ni2+ increased the incorporation of the modified nucleotide dideoxy-CMP in reactions using varying ratios of dideoxy-CTP/dCTP.

PubMed ID: 8162578 Exiting the NIEHS site

MeSH Terms: Cytidine Triphosphate/metabolism; DNA Polymerase II/drug effects; DNA Polymerase II/metabolism*; DNA Replication*; Hela Cells; Humans; Magnesium/metabolism; Nickel/metabolism*; Nickel/pharmacology

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