Biopolym. Cell. 1989; 5(4):90-97.
Cell Biology
A problem on the supramolecular organization of eukaryotic nucleoids
1Blokhin D. Yu., 1Struchkov V. A.
  1. All-Union Cancer Research Centre, Academy of Medical Sciences of the USSR
    Moscow, USSR


Properties of the nucleoids isolated from murine leukemia L1210 cells were studied using capillary viscoelastometry and sedimentation methods. Thiols (2-mercaptoethanol, dithiothreitol), pronase P, RNAse A, ethanol, sodium trichloracetate, as well as heating to 60 °C and 95 °C have been studied for their effects on nucleoids. Nucleoid cage damage by thiols or pronase has been shown to cause decompactization of the complex without loss of superhelical conformation by DNA domains. Decompactization of the complex structure is also observed when isolated nucleoids are exposed to RNAse A in the medium with low ionic strength; no decompactizing effect of RNAse A on the nucleoid structure has been revealed in other variants of enzyme treatments. Heating of the nucleoid preparations up to the "melting temperature" of the DNA double helix is shown to induce total relaxation of superhelical domains and degradation of the complex.


[1] Filippovich IV, Sorokina NI. Supercoiled DNA of the cell nucleus. Usp Sovrem Biol. 1983; 95(2):163-80.
[2] Glazkov MV. Structural-functional organization of DNA in the interphase nucleus. Structural aspects. Mol Biol (Mosk). 1988;22(1):16-30.
[3] Nicolini C. Chromatin structure: from nuclei to genes (review). Anticancer Res. 1983;3(2):63-86.
[4] Igo-Kemenes T, Zachau HG. Domains in chromatin structure. Cold Spring Harb Symp Quant Biol. 1978;42 Pt 1:109-18.
[5] Zbarsky IB. Protein comPosition and organization of the nuclear matrix. BioPolym Cell. 1985; 1(1):26-32.
[6] Igo-Kemenes T, Horz W, Zachau HG. Chromatin. Annu Rev Biochem. 1982;51:89-121.
[7] Cook PR, Brazell IA. Conformational constraints in nuclear DNA. J Cell Sci. 1976;22(2):287-302.
[8] Benyajati C, Worcel A. Isolation, characterization, and structure of the folded interphase genome of Drosophila melanogaster. Cell. 1976;9(3):393-407.
[9] Cook PR, Brazell IA, Jost E. Characterization of nuclear structures containing superhelical DNA. J Cell Sci. 1976;22(2):303-24.
[10] Cook PR, Brazell IA. The superhelical density of nuclear DNA from human cells. Eur J Biochem. 1977;74(3):527-31.
[11] Blokhin DYu, Struchkov VA. CaPillary viscoelastometry of eukaryotic nucleoids. Biopolym. Cell. 1988; 4(4):203-11.
[12] Blokhin DIu. Damage to the supramolecular DNA complex in the mechanism of action of antitumor drugs. Vopr Onkol. 1985;31(11):60-6.
[13] Blokhin DYu. Nucleoids of tumor cells: structural organization, nauk. Moscow, 1986; 23 P.
[14] Volodin YuYu, Bergolts VV, Blokhin DYu. Processing sedimentation analysis using an automated system based on a microcomPuter. Khim Farm Zh. 1988; 22(11):1381-1385.
[15] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[16] Andoh T, Ide T. A novel DNA-protein complex in cultured mouse fibroblasts, strain L-P3. Exp Cell Res. 1972;73(1):122-8.
[17] Ando T, Ide T. Disulfide bridges in proteins linking DNA in cultured mouse fibroblasts, strain L-P3. Exp Cell Res. 1972;74(2):525-31.
[18] Hartwig M. Organization of mammalian chromosomal DNA: supercoiled and folded circular DNA subunits from interphase cell nuclei. Acta Biol Med Ger. 1978;37(3):421-32.
[19] Pinon R, Salts Y. Isolation of folded chromosomes from the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1977;74(7):2850-4.
[20] Bresler SE, Noskin LA, Suslov AV. Changes in the induction and repair of double-stranded DNA breaks in pro- and eukaryote cells. I. Use of a Zimm elastoviscosimeter to study induction of double-stranded DNA breaks in gamma-irradiated Escherichia coli cells. Mol Biol (Mosk). 1980;14(6):1289-1300.
[21] Nechayevskiy YuV, Strazhevskaya NB. Gamma-induced damage of sperm loach supercoiled DNA. DNA damage and rePair. Ed. AI Gaziev. Pushchino, 1980; 107-114.
[22] Ide T, Nakane M, Anzai K, Ando T. Supercoiled DNA folded by non-histone proteins in cultured mammalian cells. Nature. 1975;258(5534):445-7.
[23] Yukhas PP, Sirota NP, Gaziyev AI.The study of proteins associated with DNA Ehrlich ascites carcinoma cells. DNA damage and repair. Ed. AI Gaziev. Pushchino, 1980; 190-202.
[24] Blokhin DIu. Localization of protein zones in polyacrylamide gel using the silver staining method. Lab Delo. 1988;(8):30-3.
[25] Paponov VD, Sigora GA, Rad'ko SP. Structural heterogeneity of chromatin preparations at the level of DNA topology. Biull Eksp Biol Med. 1987;104(11):557-9.
[26] Razin SV, Chernokhvostov VV, Rudin AV, Zbarskii IB, Georgiev GP. Proteins tightly bound to DNA at sites of its attachment to the interphase nuclear matrix. Dokl Akad Nauk SSSR. 1982;263(4):1019-21.
[27] Zbarskii IB. Structural organization and functional role of nuclear matrix. Structural and functional asPects of DNA replication and rePair: Proc. All-Union symp. Pushchino, 1983; 3-21.
[28] Lange CS. The organization and repair of mammalian DNA. FEBS Lett. 1974;44(2):153-6.
[29] Welsh RS, Vyska K. Organization of highly purified calf thymus DNA. I. Cleavage into subunits and release of phosphopeptides. Biochim Biophys Acta. 1981;655(3):291-306.
[30] Alesenko AV, Pantaz EA. Differences in the composition of nuclear and chromatin phospholipids in proliferating rat liver cells after partial hepatectomy. Biokhimiia. 1983;48(2):263-8.
[31] Chernokhvostov VV. The nuclear matrix of eukaryotic cells: some questions of isolation, structure and functioning. Usp Sovrem Biol. 1985; 99(3): 371-384.
[32] Engelhardt P, Plagens U, Zbarsky IB, Filatova LS. Granules 25-30 nm in diameter: basic constituent of the nuclear matrix, chromosome scaffold, and nuclear envelope. Proc Natl Acad Sci U S A. 1982;79(22):6937-40.
[33] Cook PR, Brazell IA. Spectrofluorometric measurement of the binding of ethidium to superhelical DNA from cell nuclei. Eur J Biochem. 1978;84(2):465-77.