Biopolym. Cell. 1993; 9(5):54-61.
http://dx.doi.org/10.7124/bc.000372
Structure and Function of Biopolymers
The way of defence of cyanophage LPP-3 DNA against restriction-modification systems in the cells cyanobacterium Plectonema boryanum
1Mendzhul M. I., 1Syrchin S. A., 1Averkiev A. A., 2Rebentish B. A.
  1. D. K. Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
    154, Academika Zabolotnogo Str., Kyiv, Ukraine, 03680
  2. Research Institute for Genetics and Selection of Industrial Microorganisms
    1-st Dorozhniy pr., 1, Moscow, Russian Federation, 117545

Abstract

HPLC method was used to study some peculiarities of cyanobacterium Plecloneina boryanum and cyanophage LPP-3 DNA composition. The elution profile analysis of the products of DNA acid hydrolysis enable to identify in DNA P. boryanum apart of canonical bases the presence of 4.5 % N-6-melhyladenine and 1.2 % 5-methyl-cytosine, DNA LPP-3 have 0,8 % 5-methyIcytosine and no N-6-methyladenine. The presence of 5-methylcytosine was detected only due to the application of the modified hydrolysis method HF. The restriction endonucleases-isoshisomers, DNA hydrolysis of which depend on presence of methylated bases in the recognition sites, were used to detect site-specific methylation. The comparison of the products of the DNA LPP-3 and P. boryanum fermentative hydrolysis by Mspl and Hpall; Sau3A, Mbol and Dpnl; Apyl and Mval restrictases enable to determined that DNA LPP-3 and P. boryanum has a high methylation degree of the inner cytosine in the CC(A/T)GG site and P. boryanum on the adenine in the GATC site; CCGG site wasn't methylated in both DNA. Proceeding from the data obtained a conclusion can be drown on the presence of dam- and dcml-like restriction–modification systems in P. boryanum. Defence of viral DNA against host R-M systems occur by cytosine methylation in sequence CmC(A/T)GG and counter-selection BamHI sites.
Full text: (PDF, in Russian)

References

[1] Alexandrushkina NI, Kirnos MD, Magradze NM, Zamchuk LA, Goldfarb DM, Vanyushin BF. A cytosine analog in Shigella sonnei “UFA” phage DNA. Russian Journal of Bioorganic Chemistry. 1978; 4(9):1191-6
[2] Kirnos MD, Aleksandrushkina NI, Vaniushin BF. 5-Methylcytosine in pyrimidine sequences of plant and animal DNA: specificity of methylation. Biokhimiia. 1981;46(8):1458-74.
[3] Lysenko TG, Shainskaya OA, Gerashchenko IV, Syrchin SA. Comparison of two methods for obtaining cell-free extracts of cyanobacteria for studying the activity of dehydrogenases. Mikrobiol Zh. 1989; 51(3):87-91.
[4] Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab, 1982; 545 p.
[5] Mendzhul MI, Syrchin SA, Averkiev AA, Busakhina IV. Comparison of different methods of cyanophage LPP-3 isolation and its DNA. Mikrobiol Zh. 1992; 54(2):70-4.
[6] Mendzhul MI, Syrchin SA, Rebentish BA, Averkiev AA, Busakhina IV. The resistance of the DNA of cyanophage LPP-3 to the action of different restriction endonucleases. Mikrobiol Z. 1993;55(4):47-53.
[7] Nesterova NV, Kalinichenko TS, Melnik AI, Syrchin SA. Degradation of cyanobacteria genome with reproduction of some cyanophages. Mikrobiol Zh. 1991; 53(3):43-9.
[8] Nesterova NV, Sagun TS, Pilipenko VG. Identificatiom cyanophage LPP-3. V Congress Ukrain. Soc.: Proc. of reports. Kiev: Naukova Dumka, 1974: 163-5.
[9] Nesterova NV, Sagun TS, Spivak NYa, Votselko SK. Characteristic of cyanophage LPP-3A gemone. Mikrobiol Z. 1982; 44(3): 34-8.
[10] Adams DG. Isolation and restriction analysis of DNA from heterocysts and vegetative cells of cyanobacteria. J Gen Microbiol. 1988;134(11):2943-9.
[11] Bancroft I, Smith RJ. An analysis of restriction endonuclease sites in cyanophages infecting the heterocystous cyanobacteria Anabaena and Nostoc. J Gen Virol. 1988;69 ( Pt 3):739-43.
[12] Brown PTH. DNA methylation in plants and its role in tissue culture. Genome. 1989;31(2):717–29.
[13] Catania J, Keenan BC, Margison GP, Fairweather DS. Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid. Anal Biochem. 1987;167(2):347-51.
[14] Chirikjian JG. Gene amplification and analysis. Restriction endonucleases and methylases. New York; Amsterdam; London : Elsevier, 1987. Vol. 5. 303 p.
[15] Doolittle MM, Sirotkin K. Bacteriophage T2 and T4, dam+ and damh and Eco dam+ methylation: preference at different sites. Biochim Biophys Acta. 1988;949(2):240-6.
[16] Hahn DR, McHenney MA, Baltz RH. Characterization of FP22, a large streptomycete bacteriophage with DNA insensitive to cleavage by many restriction enzymes. J Gen Microbiol. 1990;136(12):2395-404.
[17] Kaneko T, Katoh K, Fujimoto M, Kumagai M, Tamaoka J, Katayama-Fujimura Y. Determination of the nucleotide composition of a deoxyribonucleic acid by high-performance liquid chromatography of its enzymatic hydrlysate: a review. J Microbiol Methods. 1986;4(5-6):229–40.
[18] Krüger DH, Bickle TA. Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts. Microbiol Rev. 1983;47(3):345-60.
[19] Lambert GR, Carr NG. Resistance of DNA from filamentous and unicellular cyanobacteria to restriction endonuclease cleavage. Biochim Biophys Acta. 1984;781(1-2):45-55.
[20] McClelland M. Selection against dam methylation sites in the genomes of DNA of enterobacteriophages. J Mol Evol. 1984-1985;21(4):317-22.
[21] Padhy RN, Hottat FG, Coene MM, Hoet PP. Restriction analysis and quantitative estimation of methylated bases of filamentous and unicellular cyanobacterial DNAs. J Bacteriol. 1988;170(4):1934-9.
[22] Pogany G, Jeney A, Major J, Lapis K. Sugar-nucleotides in 5-hexyl-2-deoxyuridinetreated lewis lung tumor cells by HPLC. Chromatogram. 1989; 10(3):2-3.
[23] Roberts RJ. Restriction enzymes and their isoschizomers. Nucleic Acids Res. 1988;16 Suppl:r271-313.
[24] Schroeder C, Jurkschat H, Meisel A, Reich JG, Krüger D. Unusual occurrence of EcoP1 and EcoP15 recognition sites and counterselection of type II methylation and restriction sequences in bacteriophage T7 DNA. Gene. 1986;45(1):77-86.
[25] Sekulic S, Haddad PR. Effects of peak tailing on computer optimisation procedures for high-performance liquid chromatography. J Chromatogr.1988;459:65–77.
[26] Singhal RP, Landes JP. High-performance liquid chromatographic analysis of DNA composition and DNA modification by chloroacetaldehyde. J Chromatogr. 1988;458:117-28.