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Interaction features of adenine DNA glycosylase MutY from E. coli with DNA substrates

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Abstract

Kinetic characteristics of specific recognition of damaged base by the DNA glycosylase MutY in model DNA substrates, containing oxoG/A-, G/A-, oxoG/C- and F/G pairs in the central position, were investigated. Conformational changes of the MutY enzyme during the recognition of the damaged base in DNA have been recorded by the change in the fluorescence intensity of tryptophan residues using the stopped-flow technique in real time. DNA duplexes containing a fluorescein residue were used for the registration of DNA conformational changes. Analysis of the kinetic curves allowed us to determine the values of rate constants for the kinetic stages of the interaction. It was shown that nonspecific contacts between the DNA-binding site of the enzyme and the DNA duplex are formed at the first stage of the interaction. It was found that the discrimination of Gua and oxoGua bases occurs at the second stage of the MutY interaction with the DNA duplex. The data obtained for the oxoG/C-substrate showed that the recognition of the base located opposite oxoGua also occurs at this stage.

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Abbreviations

MutY:

adenine DNA glycosylase

BER:

base excision repair

oxoGua:

7,8-dihydro-8-oxoguanine

AP:

apurinic/apyrimidinic site

F:

(2R,3S)-2-(hydroxymethyl)-3-hydroxytetrahydrofuran residue

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Authors and Affiliations

  1. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia

    T. E. Tyugashev, A. A. Kuznetsova, N. A. Kuznetsov & O. S. Fedorova

  2. Novosibirsk State University, Novosibirsk, 630090, Russia

    T. E. Tyugashev, A. A. Kuznetsova, N. A. Kuznetsov & O. S. Fedorova

Authors
  1. T. E. Tyugashev
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  2. A. A. Kuznetsova
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  3. N. A. Kuznetsov
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  4. O. S. Fedorova
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Corresponding authors

Correspondence to N. A. Kuznetsov or O. S. Fedorova.

Additional information

Original Russian Text © T.E. Tyugashev, A.A. Kuznetsova, N.A. Kuznetsov, O.S. Fedorova, 2017, published in Bioorganicheskaya Khimiya, 2017, Vol. 43, No. 1, pp. 18–28.

The paper is based on the materials of the “Chemical Biology 2016” conference; Novosibirsk, Russia, July 24–29, 2016.

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Tyugashev, T.E., Kuznetsova, A.A., Kuznetsov, N.A. et al. Interaction features of adenine DNA glycosylase MutY from E. coli with DNA substrates. Russ J Bioorg Chem 43, 13–22 (2017). https://doi.org/10.1134/S1068162017010101

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  • DOI: https://doi.org/10.1134/S1068162017010101

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