Skip to main content
SpringerLink
Log in

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

  • Published:
Biochemistry (Moscow) Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

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

References

  1. Kobayashi, I. (2001) Nucleic Acids Res., 29, 3742–3756.

    Article  PubMed  CAS  Google Scholar 

  2. Tao, T., Bourne, J. C., and Blumenthal, R. M. (1991) J. Bacteriol., 173, 1367–1375.

    PubMed  CAS  Google Scholar 

  3. Som, S., and Freidman, S. (1994) Nucleic Acids Res., 24, 5347–5353.

    Article  Google Scholar 

  4. Som, S., and Friedman, S. (1997) J. Bacteriol., 179, 964–967.

    PubMed  CAS  Google Scholar 

  5. Karyagina, A., Shilov, I., Tashlitskii, V., Khodoun, M., Vasil’ev, S., Lau, P. C., and Nikolskaya, I. (1997) Nucleic Acids Res., 25, 2114–2120.

    Article  PubMed  CAS  Google Scholar 

  6. Shilov, I., Tashlitskii, V., Khodoun, M., Vasil’ev, S., Alekseev, Ya., Kuzubov, A., Kubareva, E., and Karyagina, A. (1998) Nucleic Acids Res., 26, 2659–2664.

    Article  PubMed  CAS  Google Scholar 

  7. Vorob’eva, O. V., Vasil’ev, S. A., Karyagina, A. S., Oretskaya, T. S., and Kubareva, E. A. (2000) Mol. Biol. (Moscow), 34, 1074–1080.

    CAS  Google Scholar 

  8. Nikol’skaya, I. I., Kartashova, I. M., and Lopatina, N. G. (1983) Mol. Genet. (Moscow), 12, 5–10.

    Google Scholar 

  9. Denjmuhametov, M. M., Zakharova, M. V., Kravecz, A. N., Pertsev, A. V., Sineva, E. V., Repyk, A. V., Beletskaya, I. V., Gromova, E. S., and Solonin, A. S. (1997) Mol. Biol. (Moscow), 31, 831–838.

    Google Scholar 

  10. Denjmuhametov, M. M., Brevnov, M. G., Zakharova, M. V., Repyk, A. V., Solonin, A. S., Petrauskene, O. V., and Gromova, E. S. (1998) FEBS Lett., 433, 233–236.

    Article  Google Scholar 

  11. Kunkel, T. (1985) Proc. Natl. Acad. Sci. USA, 82, 488–492.

    Article  PubMed  CAS  Google Scholar 

  12. Varfalomeev, S. D., and Gurevich, K. G. (1998) Biokinetics [in Russian], Fair Press, Moscow.

    Google Scholar 

  13. Karyagina, A. S., Alexeevski, A. V., Golovin, A. V., Spirin, S. A., Vorob’eva, O. V., and Kubareva, E. A. (2003) Biophysics (Moscow), 48,Suppl. 1, 45–55.

    Google Scholar 

  14. Fatemi, M., Hermann, A., Pradhan, S., and Jeltsch, A. (2001) J. Mol. Biol., 309, 1189–1199.

    Article  PubMed  CAS  Google Scholar 

  15. Streeter, S. D., Papapanagiotou, I., McGeehan, J. E., and Kneale, G. G. (2004) Nucleic Acids Res., 32, 6445–6453.

    Article  PubMed  CAS  Google Scholar 

  16. Karyagina, A. S., Lunin, V. G., Levtechenko, I. Ya., Labbe, D., Brousseau, R., Lau, P. C., and Nikolskaya, I. I. (1995) Gene, 157, 93–96.

    Article  PubMed  CAS  Google Scholar 

  17. Kubareva, E. A., Walter, J., Karyagina, A. S., Vorob’eva, O. V., Lau, P. C. K., and Trautner, T. (2002) BioTechniques, 33, 526–531.

    PubMed  CAS  Google Scholar 

  18. Vorob’eva, O. V. (2004) (Cytosine-5)-DNA Methyltrans-ferase SsoII as Bifunctional Protein: Studying Interaction with Methylation Site and with Promoter Gene Region of SsoII Restriction-Modification System: Candidate’s dissertation [in Russian], Moscow State University, Moscow.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Chemical Faculty, Lomonosov Moscow State University, 119991, Moscow, Russia

    E. A. Fedotova & T. S. Oretskaya

  2. Pushchino State University, pr. Nauki 3, 142290, Pushchino, Moscow Region, Russia

    A. S. Protsenko

  3. Skryabin Institute of Biochemistry and Physiology of Microorganisms, pr. Nauki 5, 142290, Pushchino, Moscow Region, Russia

    A. S. Protsenko, M. V. Zakharova & A. S. Solonin

  4. Gamaleya Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, ul. Gamalei 18, 123098, Moscow, Russia

    N. V. Lavrova & A. S. Karyagina

  5. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    A. V. Alekseevsky & E. A. Kubareva

Authors
  1. E. A. Fedotova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. S. Protsenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. V. Zakharova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. N. V. Lavrova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. V. Alekseevsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T. S. Oretskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. S. Karyagina
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A. S. Solonin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. E. A. Kubareva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E. A. Kubareva.

Additional information

Original Russian Text © 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. Kubareva, 2009, published in Biokhimiya, 2009, Vol. 74, No. 1, pp. 108–116.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM08-185, December 14, 2008.

Prefix “d” (deoxy) in oligodeoxyribonu-cleotide and DNA duplex names is skipped

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fedotova, E.A., Protsenko, A.S., Zakharova, M.V. et al. SsoII-like DNA-methyltransferase Ecl18kI: Interaction between regulatory and methylating functions. Biochemistry Moscow 74, 85–91 (2009). https://doi.org/10.1134/S0006297909010131

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0006297909010131

Key words

Search

Navigation