Resolution of DNA polymerase-α-primase complex and primase free DNA polymerase α from embryonic chicken brain.
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Date
1987-03
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Abstract
DNA polymerase-α from embryonic chicken brain was resolved on DEAEcellulose
into 3 component activities that remained distinct upon rechromatography.
Product formation by each activity required exogenously added template-primer DNA, all 4
deoxynucleoside triphosphates, and a divalent metal cation. Each form incorporated [3H]-
dTTP or [3H]-dCTP into a high molecular weight product that was identified as DNA by
its chromatographic behavior and its sensitivity to DNase. High ionic strength, Nethylmaleimide,
and the polymerase-α-specific inhibitor aphidicolin inhibited each activity;
the apparent Ki, value of aphidicolin was 3·0 μΜ in each case. Based on these results, the 3
activities were identified as multiple forms of DNA polymerase-α . Experiments using
embryonic chicken brains of various ages indicated that polymerase-α1, and polymerase-α3
reached maximal activity in 9-day-old embryos, while polymerase-α2 activity was elevated
at a slightly later developmental stage. Using poly (dC) as template, high primase activity
was detected in polymerase-α1, fractions.
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Keywords
DNA polymerase-α, aphidicolin inhibition, primase, N-ethylmaleimide inhibition, DNA chain initiation
Citation
Simet Ira, Ray Satyajit, Basu Subhash. Resolution of DNA polymerase-α-primase complex and primase free DNA polymerase α from embryonic chicken brain. Journal of Biosciences. 1987 Mar; 11(1-4): 361-378.