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Nucleic Acids Research, 1983, Vol. 11, No. 1 193-202
© 1983

ENZYMOLOGY

Subspecies of DNA polymerase {alpha} from calf thymus with different fidelity in copying synthetic template-primers

Sabine Brosius1, Frank Grosse and Gerhard Krauss*

Zentrum Biochemie, Abteilung Biophysikalische Chemie, Medizinische Hochschule Hannover, Konstanty-Gutschnow Strasse 8 3000 Hannover, FRG

Received November 16, 1982. Accepted December 4, 1982.

Three different subspecies of DNA polymerase a from calf thymus sedimenting at 9 S, 7 S and 5.7 S have been investigated with respect to their accuracy of in vitro DNA synthesis on poly(dA) (dT)16 and poly d(AT) as template-primers. Our results indicate that the structure of DNA polymerase {alpha} has a strong influence on the accuracy of DNA synthesis. The 9 S enzyme shows a mis-incorporation frequency of about 1:100 OOO. An error rate of 1:15 OOO is measured for the 7 S species. The 5.7 S enzyme for which an error rate of 1:3 OOO is determined, has to be considered as error prone. Lowering the rate of DNA synthesis leads to a decrease in fidelity. The single stranded DNA binding protein from E.coli increases the accuracy of the 5.7 S and the 7 S enzyme by a factor of two. Mn2+ decreases the fidelity of all three subspecies in a concentration dependent manner.

1Present address: Institut füur Mikrobiologie, Ruhniniversität Bochum, D-4630 Bochum, FRG

*To whom reprint requests should be sent


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