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Nucleic Acids Research, 1974, Vol. 1, No. 10 1233-1248
© 1974

Articles

Deoxyribonucleic acid of Cancer pagurus. II. Template activity for a DNA-dependent DNA polymerase of eukaryotic cells

Anne-Marie de Recondo1, Danielle Londos-Gagliardi2 and Genevieve Aubel-Sadron2

1Institut de Recherches Scientifiques sur le Cancer BP No.8, 94800-Villejuif, France 2Centre de Biophysique Moléculaire CNRS, 4502-Orléans, France

Received July 12, 1974. The template activity of Cancer pagurus DNA and its two components (poly d(A–T) and main component) in response to a DNA polymerase purified from regenerating rat liver has been studied and compared to the results previously obtained with synthetic templates. In the double-stranded native state, whole crab DNA and the main component were poor templates. Their replication was increased by thermal denaturation and inhibited by actinomycin. Like the synthetic copolymer poly[d(A–T).d(T–A)], native crab poly d(A–T) could be copied and its duplication was not inhibited by actinomycin. The structural difference between native poly d(A–T) Form I, isolated on a density gradient, and partially renatured poly d(A–T) Form II, isolated on hydroxylapatite, resulted in a modification of their template activity. The kinetic studies of [3H]dGMP and [3H] dAMP incorporation confirmed the importance of single-stranded regions (particulary dC regions) in the initiation of the in vitro duplication.


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