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Nucleic Acids Research, 1985, Vol. 13, No. 15 5685-5695
© 1985

Articles

On the fidelity of DNA polymerase {alpha}: the influence of {alpha}-thio dNTPs, Mn2+ and mismatch repair

Axel Rienitz1, Frank Grosse2, Helmut Blöcker3, Ronald Frank3 and Gerhard Krauss*

Laboratorium für Biochemie, Universtät Bayreuth Postfach 3008, D-8580 Bayreuth 1Zentrum für Molekularbiologie Heidelberg 2Max-Planck Institut für Experimentelle Medizin Göttingen 3Gesellschaft für Biotechnologische Forschung mbH Braunschweig, FRG

*To Whom Correspondence should be addressed

Received May 28, 1985. Revised July 19, 1985. Accepted July 19, 1985.

The FE X174aml6 revertant system has been used to investigate the influence of {alpha} -thio-dNTPs and of Mn2+ on the fidelity of the 9S DNA polymerase {alpha} from calf thymus. Upon substituting dGTP by {alpha}-thio-dGTP during the in vitro replication, a nearly tenfold decrease in the frequency of G: G and G: T mispairs is observed. The formation of all other mispairs is not changed in the presence of the corresponding {alpha}-thio-dNTP. Mn2+ at concentrations of 0.5 mM does not influence the frequencies of the mispairs. The expression rate of errors formed during in vitro replication in the (-) strand has been determined for all mispairs detectable in the FE Xaml6 system. The (-) strand expression of G: T, T: T and C: T mismatches is about 50% whereas for A: G, G: G and C: A mismatches it is clearly below 50%. We conclude that the different base-base mismatches are repaired with different efficiencies.


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