Nucleic Acids Research, 1984, Vol. 12, No. 7 3173-3183
© 1984
ENZYMOLOGY |
Template–dependent variation in the relative fidelity of DNA polymerase I of Escherichia coli in the presence of Mg2+ versus Mn2+
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine Houston, TX 77030, USA
*To whom correspondence should be addressed
Received January 3, 1984. Revised March 12, 1984. Accepted March 12, 1984.
The fidelity of E. coli DNA polymerase I in the preseace of Mg2+ vs Mn2+ was examined at many positions along natural DNA templates, by use of an electrophoretic assay of misincorporation. Although there was an overall greater tendency for misincorporation to occur in Mn2+-activated chain elongation, some specific sites on the template were more prone to misincorporation with Mg2+ and others with Mn2+. This sequence–dependent effect was seen in spite of the finding that the relative rate of incorporation of the correct nucleotide at different positions an the template was essentially the same with Mg2+ and Mn2+. In agreement with previous studies, the fidelity of E. coli pol I was higher at activating, than at inhibiting, concentrations of Mg2+. The results reveal new complexities regarding the role of divalent cation in the control of fidelity in DNA synthesis and attest to the dynamic nature of interactions between DNA polymerase, its substrates and divalent metal activator during the course of polymerization on natural templates.
+Present address: Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA