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Nucleic Acids Research, 1985, Vol. 13, No. 1 261-274
© 1985

Articles

DNA polymerase {alpha} and models for proofreading

John Abbotts and Lawrence A. Loeb+

Department of Biochemistry, and Joseph Gottstein Memorial Cancer Research Laboratory, Department of Pathology SM-30, University of Washington Seattle, WA 98195, USA

+To whom reprint requests should be addressed

Received September 21, 1984. Revised December 7, 1984. Accepted December 7, 1984.

Using a modified system to measure fidelity at an amber site in øX174, we have employed DNA polymerase {alpha} to test different mechanisms for proofreading. DNA polymerase {alpha} does not exhibit the characteristics of "kinetic proofreading" seen with procaryotic polymerases. Polymerase {alpha} shows no evidence for a "next nucleotide" effect, and added deoxynucleoside monophosphates do not alter fidelity. Pyrophosphate, which increases error rates with a procaryotic polymerase, appears to weakly improve polymerase a fidelity. DNA polymerase {alpha} does exhibit a dramatic increase in error rate in the presence of a deoxycytidine thiotriphosphate (dCTP{alpha}S), but this enhanced mutagenesis also occurs under conditions where kinetic proofreading should be otherwise defeated. This particular effect with dCTP{alpha}S appears specific for DNA polymerase {alpha} and is not seen with the other polymerases tested.


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