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Nucleic Acids Research, 1983, Vol. 11, No. 11 3753-3765
© 1983

CHEMISTRY

Uracil in deoxyribonucleotide polymers reduces their template-primer activity for E. coli DNA polymerase I

J. A. vilpo and J. Ridell

Department of Clinical Chemistry, University of Oulu SF-90220 Oulu 22, Finland

Received April 18, 1983. Accepted May 12, 1983.

The physical and biochemical properties of two pairs of synthetic DNA template–primers were investigated. The copoly–mer poly(dA–dU)·poly(dA–dU) and the homopolymer duplex polyldA) –poly(dU) were characterized by a lower Tm and by a higher buoyant density value than the respective thymine polynucleotides poly(dA–dT) –poly(dA–dT) and poly(dA) –poly(dT). The polymerizing and the primer terminus adding reactions of a homogenous E. coli DNA polymeraae I preparation, as measured by incorporation of [3H]dAMP into the acid-insoluble fraction, were significantly poorer with uracil-containing template–primers than with thymine templates. Moreover, the uracil–containing polynucleotides inhibited the polymerizing activity of DNA polymerase I to a greater extent than the thymine polynucleotides, when the enzymatic activity was investigated with a dATP/dTTP/dUTP–free incorporation system making use of poly(dl–dC) –poly(dl–dC) as the template–primer.


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