Mechanism of DNA polymerase I: Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions

V. Mizrahi; P. Benkovic; S. J. Benkovic

doi:10.1073/pnas.83.16.5769

Mechanism of DNA polymerase I: Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions

V. Mizrahi, P. Benkovic, S. J. Benkovic

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Abstract

Mechanistic features of several processes involved in the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I have been established. The exonuclease → polymerase activity switch involved in the excision/incorporation mode of idling-turnover occurs without an intervening dissociation of the enzyme from its DNA substrate. Comparative studies on the pyrophosphorolysis kinetics of related DNA substrates indicate a significant dependence of the reaction rate upon the DNA sequence within the duplex region upstream of the primer-template junction. Finally, a gel electrophoretic analysis of the products of the idling-turnover reaction has provided direct evidence for an alternative DNA sequence-dependent misincorporation/excision pathway.

Original language	English (US)
Pages (from-to)	5769-5773
Number of pages	5
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	83
Issue number	16
DOIs	https://doi.org/10.1073/pnas.83.16.5769
State	Published - 1986

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title = "Mechanism of DNA polymerase I: Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions",

abstract = "Mechanistic features of several processes involved in the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I have been established. The exonuclease → polymerase activity switch involved in the excision/incorporation mode of idling-turnover occurs without an intervening dissociation of the enzyme from its DNA substrate. Comparative studies on the pyrophosphorolysis kinetics of related DNA substrates indicate a significant dependence of the reaction rate upon the DNA sequence within the duplex region upstream of the primer-template junction. Finally, a gel electrophoretic analysis of the products of the idling-turnover reaction has provided direct evidence for an alternative DNA sequence-dependent misincorporation/excision pathway.",

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Mechanism of DNA polymerase I: Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions. / Mizrahi, V.; Benkovic, P.; Benkovic, S. J.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 83, No. 16, 1986, p. 5769-5773.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Mechanism of DNA polymerase I

T2 - Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions

AU - Mizrahi, V.

AU - Benkovic, P.

AU - Benkovic, S. J.

PY - 1986

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AB - Mechanistic features of several processes involved in the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I have been established. The exonuclease → polymerase activity switch involved in the excision/incorporation mode of idling-turnover occurs without an intervening dissociation of the enzyme from its DNA substrate. Comparative studies on the pyrophosphorolysis kinetics of related DNA substrates indicate a significant dependence of the reaction rate upon the DNA sequence within the duplex region upstream of the primer-template junction. Finally, a gel electrophoretic analysis of the products of the idling-turnover reaction has provided direct evidence for an alternative DNA sequence-dependent misincorporation/excision pathway.

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Mechanism of DNA polymerase I: Exonuclease/polymerase activity and DNA sequence dependence of pyrophosphorolysis and misincorporation reactions

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