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Novel DNA glycosylases from Mycobacterium tuberculosis

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Abstract

Oxidized bases are removed from DNA of Escherichia coli by enzymes formamidopyrimidine DNA glycosylase (Eco-Fpg) and endonuclease VIII (Eco-Nei) of the same structural family Fpg/Nei. New homologs of these enzymes not characterized earlier have been found in genomes of Actinobacteria. We have cloned and expressed two paralogs (Mtu-Nei2 and Mtu-Fpg2) from 36KAZ and KHA94 isolates of Mycobacterium tuberculosis and studied their ability to participate in DNA repair. Under heterologous expression in E. coli, Mtu-Nei2 decreased the rate of spontaneous mutagenesis in the rpoB gene, whereas Mtu-Fpg2 moderately increased it, possibly due to absence of residues crucially important for catalysis in this protein. Mtu-Nei2 was highly active toward double-stranded DNA substrates containing dihydrouracil residues and apurine-apyrimidine sites and was less efficient in cleavage of substrates containing 8-oxoguanine and uracil residues. These lesions, as well as 8-oxoadenine residues, were also recognized and removed by the enzyme from single-stranded DNA. Fpg and Nei homologs from M. tuberculosis can play an important role in protection of bacteria against genotoxic stress caused by oxidative burst in macrophages.

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Abbreviations

AP:

apurine-apyrimidine

BER:

base excision repair

DHU:

5,6-dihydrouracil

ODN:

oligodeoxyribonucleotide

8-oxoG:

7,8-dihydro-8-oxoguanine

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Authors and Affiliations

  1. Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, pr. Lavrentieva 8, 630090, Novosibirsk, Russia

    V. S. Sidorenko, M. A. Rot, M. L. Filipenko, G. A. Nevinsky & D. O. Zharkov

Authors
  1. V. S. Sidorenko
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  2. M. A. Rot
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  3. M. L. Filipenko
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  4. G. A. Nevinsky
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  5. D. O. Zharkov
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Correspondence to D. O. Zharkov.

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Original Russian Text © V. S. Sidorenko, M. A. Rot, M. L. Filipenko, G. A. Nevinsky, D. O. Zharkov, 2008, published in Biokhimiya, 2008, Vol. 73, No. 4, pp. 542–552.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-319, February 3, 2008.

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Sidorenko, V.S., Rot, M.A., Filipenko, M.L. et al. Novel DNA glycosylases from Mycobacterium tuberculosis . Biochemistry Moscow 73, 442–450 (2008). https://doi.org/10.1134/S0006297908040093

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  • DOI: https://doi.org/10.1134/S0006297908040093

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