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Antirestriction Protein ArdB (R64) Interacts with DNA

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Abstract

The antirestriction ArdB protein inhibits the endonuclease activity of type I restriction/modification (RM) systems in vivo; however, the mechanism of inhibition remains unknown. In this study, we showed that recombinant ArdB from Escherichia coli cells co-purified with DNA. When overexpressed in E. coli cells, a portion of ArdB protein formed insoluble DNA-free aggregates. Only native ArdB, but not the ArdBAD141 mutant lacking the antirestriction activity, co-purified with DNA upon anion-exchange and affinity chromatography or total DNA isolation from formaldehyde-treated cells. These observations confirm the hypothesis that ArdB blocks DNA translocation via the R subunits of the R2M2S complex of type I RM enzymes.

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Abbreviations

GST:

glutathione S-transferase

RM:

restriction/modification (system)

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Authors and Affiliations

  1. Moscow Institute of Physics and Technology, 141707, Dolgoprudny, Moscow Region, Russia

    A. A. Kudryavtseva, I. S. Okhrimenko, V. S. Didina & I. V. Manukhov

  2. State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center, 117545, Moscow, Russia

    G. B. Zavilgelsky & I. V. Manukhov

Authors
  1. A. A. Kudryavtseva
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  2. I. S. Okhrimenko
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  3. V. S. Didina
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  4. G. B. Zavilgelsky
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  5. I. V. Manukhov
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Corresponding author

Correspondence to A. A. Kudryavtseva.

Additional information

Russian Text © The Author(s), 2020, published in Biokhimiya, 2020, Vol. 85, No. 3, pp. 369–377.

Funding

The work was supported by the Russian Foundation for Basic Research (projects nos. 18-3400753 and 19-04-00495) and State Order no. 6.9899. 2017/BCh.

Conflict of interest

The authors declare no conflict of interest in financial or any other area.

Compliance with ethical standards

This article does not contain any studies involving animals or human participants performed by any of the authors.

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Kudryavtseva, A.A., Okhrimenko, I.S., Didina, V.S. et al. Antirestriction Protein ArdB (R64) Interacts with DNA. Biochemistry Moscow 85, 318–325 (2020). https://doi.org/10.1134/S0006297920030074

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  • DOI: https://doi.org/10.1134/S0006297920030074

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