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Biochemistry (Moscow)

, Volume 74, Issue 1, pp 85–91 | Cite as

SsoII-like DNA-methyltransferase Ecl18kI: Interaction between regulatory and methylating functions

  • E. A. Fedotova
  • A. S. Protsenko
  • M. V. Zakharova
  • N. V. Lavrova
  • A. V. Alekseevsky
  • T. S. Oretskaya
  • A. S. Karyagina
  • A. S. Solonin
  • E. A. KubarevaEmail author
Article

Abstract

The interaction of DNA-methyltransferase Ecl18kI (M.Ecl18kI) with a fragment of promoter region of restriction-modification system SsoII was studied. It is shown that dissociation constants of M.Ecl18kI and M.SsoII complexes with DNA ligand carrying a regulatory site previously characterized for M.SsoII have comparable values. A deletion derivative of M.Ecl18kI, Δ(72–379)Ecl18kI, representing the N-terminal protein region responsible for regulation, was obtained. It is shown that such polypeptide fragment has virtually no interaction with the regulatory site. Therefore, the existence of a region responsible for methylation is necessary for maintaining M.Ecl18kI regulatory function. The properties of methyl-transferase NlaX, which is actually a natural deletion derivative of M.Ecl18kI and M.SsoII lacking the first 70 amino acid residues and not being able to regulate gene expression of the SsoII restriction-modification system, were studied. The ability of mutant forms of M.Ecl18kI incorporating single substitutions in regions responsible for regulation and methylation to interact with both sites of DNA recognition was characterized. The data show a correlation between DNA-binding activity of two M.Ecl18kI regions-regulatory and methylating.

Key words

regulation in restriction-modification systems (cytosine-5)-DNA methyltransferase DNA-protein interactions 

Abbreviations

a. a.

amino acid residue

AdoHcy

S-adenosyl-L-homocysteine

AdoMet

S-adenosyl-L-methionine

HTH

“helix-turn-helix”

MTase

DNA-methyltransferase

N is A

G, T, or C

RE

restriction endonuclease

R-M

restriction-modification

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • E. A. Fedotova
    • 1
  • A. S. Protsenko
    • 2
    • 3
  • M. V. Zakharova
    • 3
  • N. V. Lavrova
    • 4
  • A. V. Alekseevsky
    • 5
  • T. S. Oretskaya
    • 1
  • A. S. Karyagina
    • 4
  • A. S. Solonin
    • 3
  • E. A. Kubareva
    • 5
    Email author
  1. 1.Chemical FacultyLomonosov Moscow State UniversityMoscowRussia
  2. 2.Pushchino State UniversityPushchino, Moscow RegionRussia
  3. 3.Skryabin Institute of Biochemistry and Physiology of MicroorganismsPushchino, Moscow RegionRussia
  4. 4.Gamaleya Research Institute of Epidemiology and MicrobiologyRussian Academy of Medical SciencesMoscowRussia
  5. 5.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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