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Secondary structure of SsoII-like (Cytosine-5)-DNA methyltransferases N-terminal region determined by Circular dichroism spectroscopy

  • Structural-Functional Analysis of Biopolymers and Their Complexes
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Abstract

(Cytosine-5)-DNA methyltransferase SsoII (M.SsoII) has a long N-terminal region (1–71 residues) preceding the sequence with conservative motifs, which are characteristic for all DNA methyltrans-ferases of such kind. The presence of this region provides M.SsoII capability to act as a transcription regulator in SsoII restriction-modification system. To perform its regulatory function, M.SsoII binds specifically to a 15-mer inverted repeat in the promoter region of SsoII restriction-modification system genes. In the present work, properties of the protein Δ(72-379)M.Ecl18kI are studied, which is a deletion mutant of the SsoII-like DNA-methyltransferase M.Ecl18kI and is homologous to M.SsoII N-terminal region. Δ(72-379)M.Ecl18kI capability to bind specifically a DNA duplex containing the regulatory site is demonstrated. However, such a binding takes place only in the presence of high protein excess relative to DNA, which could indicate an altered structure in the deletion mutant in comparison with the full-length M.SsoII. Circular dichroism spectroscopy demonstrated that Δ(72–379)M.Ecl18kI has a strongly pronounced secondary structure and contains 32% α-helices and 20% β-strands. Amino acid sequences alignment of M.SsoII N-terminal region and transcription factors of known spatial structure is made. An assumption is made how α-helices and β-strands are arranged in M.SsoII N-terminal region.

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Abbreviations

CD:

circular dichroism

HTH:

helix-turnhelix

M.Ecl18kI:

(cytosine-5)-DNA methyltransferase Ecl18kI

M.HaeIII:

(cytosine-5)-DNA methyltransferase HaeIII

M.HhaI:

(cytosine-5)-DNA methyltransferase HhaI

M.NlaX:

(cytosine-5)-DNA methyltransferase NlaX

M.SsoII:

(cytosine-5)-DNA methyltransferase SsoII

MTase:

methyltransferase

NMR:

nuclear magnetic resonance

PAAG:

polyacrylamide gel

R-M:

restriction-modification

RE:

restriction endonuclease

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Authors and Affiliations

  1. Faculty of Bioengineering and Bioinformatics, Chemistry Department, and A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, 119991, Russia

    A. Yu. Ryazanova, L. A. Abrosimova, A. V. Alexeevsky, A. S. Karyagina, T. S. Oretskaya & E. A. Kubareva

  2. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Moscow region Pushchino, 142290, Russia

    N. V. Molochkov

  3. Scientific Research Institute for System Studies, Russian Academy of Sciences, Moscow, 117218, Russia

    A. V. Alexeevsky

  4. N.F. Gamaleya Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, Moscow, 123098, Russia

    A. S. Karyagina

  5. G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Moscow region Pushchino, 142290, Russia

    A. S. Protsenko

  6. Institut für Biochemie, Justus-Liebig-Universität, D-35392, Giessen, Germany

    P. Friedhoff

Authors
  1. A. Yu. Ryazanova
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  2. N. V. Molochkov
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  3. L. A. Abrosimova
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  4. A. V. Alexeevsky
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  5. A. S. Karyagina
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  6. A. S. Protsenko
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  8. T. S. Oretskaya
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  9. E. A. Kubareva
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Correspondence to E. A. Kubareva.

Additional information

Original Russian Text © A.Yu. Ryazanova, N.V. Molochkov, L.A. Abrosimova, A.V. Alexeevsky, A.S. Karyagina, A.S. Protsenko, P. Friedhoff, T.S. Oretskaya, E.A. Kubareva, 2010, published in Molekulyarnaya Biologiya, 2010, Vol. 44, No. 5, pp. 911–921.

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Ryazanova, A.Y., Molochkov, N.V., Abrosimova, L.A. et al. Secondary structure of SsoII-like (Cytosine-5)-DNA methyltransferases N-terminal region determined by Circular dichroism spectroscopy. Mol Biol 44, 807–816 (2010). https://doi.org/10.1134/S0026893310050183

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  • DOI: https://doi.org/10.1134/S0026893310050183

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