Abstract
The ArdA and Ocr antirestriction proteins, whose genes are in transmissible plasmids (ardA) and bacteriophage genomes (0.3 (ocr)), specifically inhibit type I restriction-modification enzymes. The Ocr protein (T7 bacteriophage) was shown to inhibit both restriction (endonuclease) and modification (methylase) activities of the EcoKI enzyme in a broad range of intracellular concentrations (starting from 10–20 molecules per cell). In contrast to Ocr, the ArdA protein (ColIb-P9 transmissible plasmid) inhibited both of the EcoKI activities only at high intracellular concentrations (30000–40000 molecules per cell). When the ArdA concentration was several fold lower, only endonuclease activity of EcoKI was inhibited. It was assumed that a poorer ArdA ability to inhibit EcoKI modification activity is related to the substantial difference in life cycle between transmissible plasmids (symbiosis with the bacterial cell) and bacteriophages (infection and lysis of bacteria). The Ocr and ArdA mutants that inhibited exclusively endonuclease activity of EcoKI were obtained. Antirestriction proteins incapable of homodimerization were assumed to inhibit only endonuclease activity of type I restriction-modification enzymes.
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Zavilgelsky, G.B., Belogurov, A.A., Mershavka, V.Yu., and Yusifov, T.N., Alleviation of Type I Restriction in the Presence of pKM101 Plasmid (incN), Mol. Biol., (Moscow), 1984, vol. 18, pp. 1590–1596.
Belogurov, A.A., Yussifov, T.N., Kotova, V.Yu., and Zavilgelsky, G.B., The Novel Gene ard of Plasmid pKM101; Alleviation of EcoKI Restriction, Mol. Gen. Genet., 1985, vol. 198, pp. 509–513.
Kotova, V.Yu., Belogurov, A.A., and Zavilgelsky, G.B., Alleviation of the Type I Restriction in the Presence of Plasmid from IncI Group: General Characteristic and Molecular Cloning of the Gene ard, Mol. Biol., (Moscow), 1988, vol. 22, pp. 270–276.
Delver, E.P., Kotova, V.Yu., Zavilgelsky, G.B., and Belogurov, A.A., Nucleotide Sequence of the Gene (ard) Encoding the Antirestriction Protein of Plasmid ColIb-P9, J. Bacteriol., 1991, vol. 173, pp. 5887–5892.
Zavilgelsky, G.B., Bakalova, T.L., Duzhii, D.E., and Kotova, V.Yu., Gene ard Coding for a Type I Restriction Inhibitor is Present in Conjugative Plasmids of Incompatibility Groups, Russ. J. Genet., 1994, vol. 30, no. 12, pp. 1582–1586.
Chilley, P.M. and Wilkins, B.M., Distribution of the ard A Family of Antirestriction Genes on Conjugative Plasmids, Microbiology, 1995, vol. 141, pp. 2157–2164.
Zavilgelsky, G.B., Antirestriction, Mol. Biol., (Moscow), 2000, vol. 34, pp. 854–862.
Dunn, J.J., Elzinga, M., Mark, K.K., and Studier, F.W., Amino Acid Sequence of the Gene 0.3 Protein of Bacteriophage T7 and Nucleotide Sequence of Its Messenger RNA, J. Biol. Chem., 1981, vol. 256, pp. 2579–2585.
Kruger, D.H., Hansen, S., and Reuter, M., The ocr Gene Function of Bacteriophages T3 and T7 Counteracts the Salmonella typhimurium DNA Restriction Systems SA and SB, J. Virol., 1983, vol. 45, pp. 1147–1149.
Bandyopadhay, P.K., Studier, F.W., Hamilton, D.L., and Yuan, R., Inhibition of the Type I Restriction-Modification Enzymes EcoB and EcoK by the Gene 0.3 Protein of Bacteriophage T7, J. Mol. Biol., 1985, vol. 182, pp. 567–578.
Walkinshaw, M.D., Taylor, P., Sturrock, S.S., et al., Structure of Ocr from Bacteriophage T7: A Protein That Mimics B-Form DNA, Mol. Cell, 2002, vol. 9, pp. 187–194.
Atanasiu, C., Su, T.-J., Sturrocks, S.S., and Dryden, D.T.F., Interaction of the ocr Gene 0.3 Protein of Bacteriophage T7 with EcoKI Restriction/Modification Enzyme, Nucleic Acids Res., 2002, vol. 30, pp. 3936–3944.
McMahon, S.A., Roberts, G.A., Johnson, K.A., et al., Extensive DNA Mimicry by the ArdA Anti-Restriction Protein and Its Role in the Spread of Antibiotic Resistance, Nucleic Acids Res., 2009, vol. 37, pp. 4887–4897.
Althorpe, N.J., Chilley, P.M., Thomas, A.T., et al., Transient Transcriptional Activation of the IncI1 Plasmid Antirestriction Gene (ardA) and SOS Inhibition Gene (psiB) Early in Conjugating Recipient Bacteria, Mol. Microbiol., 1999, vol. 31, pp. 133–142.
Murray, N.E., Type I Restriction Systems: Sophisticated Molecular Machines (a Legacy of Bertani and Weigle), Microbiol. Mol. Biol. Rev., 2000, vol. 64, pp. 412–434.
Tock, M.R. and Dryden, D.T., The Biology of Restriction and Antirestriction, Curr. Opin. Microbiol., 2005, vol. 8, pp. 466–472.
Rastorguev, S.M. and Zavilgelsky, G.B., Role of “Antirestriction” Motif in Functional Activity of Antirestriction Protein ArdA pKM101 (IncN), Russ. J. Genet., 2003, vol. 39, no. 2, pp. 218–224.
Zavilgelsky, G.B., Letuchaya, E.A., and Rastorguev, S.M., Antirestriction and Antimodification Activities of the ArdA Protein Encoded by the IncI1 Transmissive Plasmids R-64 and ColIb-P9, Russ. J. Genet., 2006, vol. 42, no. 3, pp. 333–338.
Zavilgelsky, G.B., Kotova, V.Yu., and Rastorguev, S.M., Comparative Analysis of Antirestriction Activity of ArdA (ColIb-P9) and Ocr (T7) Proteins, Biochemistry (Moscow), 2008, vol. 73, pp. 1124–1130.
Miller, J.H., Experiments in Molecular Genetics, Cold Spring Harbor: Cold Spring Harbor Laboratories, 1972.
Lutz, R. and Bujard, H., Independent and Tight Regulation of Transcriptional Units in Escherichia coli via the LacR/O, TheTetR/O and AraCI1-I2 Regulatory Elements, Nucleic Acids Res., 1997, vol. 25, pp. 1203–1210.
Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Lab., 1989, 2ed.
Sanger, F., Nicklen, S., and Coulson, A.R., DNA Sequencing with Chain-Terminating Inhibition, Proc. Natl. Acad. Sci. USA, 1977, vol. 74, pp. 5463–5467.
Landt, O., Grunert, H.P., and Hahn, U., A General Method for Rapid Site-Directed Mutagenesis Using the Polymerase Chain Reaction, Gene, 1990, vol. 96, pp. 125–128.
Thomas, A.T., Brammar, W.J., and Wilkins, B.M., Plasmid R16 ArdA Protein Preferentially Targets Restriction Activity of the Type I Restriction-Modification System, J. Bacteriol., 2003, vol. 185, pp. 2022–2025.
Zavilgelsky, G.B., Letuchaya, T.A., and Rastorguev, S.M., Antirestriction Activity of the ArdA Protein Encoded by Transmissive Plasmids R64, Mol. Biol., 2004, vol. 38, pp. 904–907.
Zavilgelsky, G.B. and Rastorguev, S.M., Antirestriction Proteins ArdA and Ocr as Efficient Inhibitors of Type I Restriction-Modification Enzymes, Mol. Biol. (Moscow), 2009, vol. 43, pp. 264–273.
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Original Russian Text © G.B. Zavilgelsky, V.Yu. Kotova, S.M. Rastorguev, 2011, published in Genetika, 2011, Vol. 47, No. 2, pp. 159–167.
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Zavilgelsky, G.B., Kotova, V.Y. & Rastorguev, S.M. Antimodification activity of the ArdA and Ocr proteins. Russ J Genet 47, 139–146 (2011). https://doi.org/10.1134/S1022795410081034
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DOI: https://doi.org/10.1134/S1022795410081034