Mechanism of the idling-turnover reaction of the large (Klenow) fragment of Escherichia coli DNA polymerase I

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

Research output: Contribution to journalArticle

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Abstract

The mechanism of the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I has been investigated. The reaction cycle involved is one of excision/incorporation, in which the 3' deoxynucleotide residue of the primer DNA strand is partitioned into its 5'-mono- and 5'-triphosphate derivatives, respectively. Mechanistic studies suggest the 5'-monophosphate product is formed in the first step by simple 3'→5' exonucleolytic cleavage. Rapid polymerization follows with the concomitant release of inorganic pyrophosphate. In the second step, the 5'-triphosphate product is generated by a pyrophosphorolysis reaction, which, despite the low concentration of pyrophosphate that has accumulated, occurs at a rate that is comparable with that of the parallel 3'→5' hydrolysis reaction.

Original languageEnglish (US)
Pages (from-to)231-235
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number2
StatePublished - Jan 1 1986

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DNA Polymerase I
Escherichia coli
DNA Primers
Polymerization
Hydrolysis
diphosphoric acid
triphosphoric acid

All Science Journal Classification (ASJC) codes

  • General

Cite this

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abstract = "The mechanism of the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I has been investigated. The reaction cycle involved is one of excision/incorporation, in which the 3' deoxynucleotide residue of the primer DNA strand is partitioned into its 5'-mono- and 5'-triphosphate derivatives, respectively. Mechanistic studies suggest the 5'-monophosphate product is formed in the first step by simple 3'→5' exonucleolytic cleavage. Rapid polymerization follows with the concomitant release of inorganic pyrophosphate. In the second step, the 5'-triphosphate product is generated by a pyrophosphorolysis reaction, which, despite the low concentration of pyrophosphate that has accumulated, occurs at a rate that is comparable with that of the parallel 3'→5' hydrolysis reaction.",
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Mechanism of the idling-turnover reaction of the large (Klenow) fragment of Escherichia coli DNA polymerase I. / Mizrahi, V.; Benkovic, P. A.; Benkovic, S. J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 83, No. 2, 01.01.1986, p. 231-235.

Research output: Contribution to journalArticle

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