Biopolym. Cell. 1995; 11(3-4):66-72.
The formation of high molecular weight dna fragments: the hallmark of apoptosis or early genomic events in stress response?
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
Abstract
We studied the peculiarities of nuclear DNA fragmentation in primary culture of marine thymocytes and in human lymphoblastoid cell culture (line CEM) induced to apoptosis by various influences. We showed that in apoptotic cells the ordered high molecular weight DNA cleavage, recognizable as DNA fragment release of 50–100 and 250–300 kb in length, preceded to typical nuclear DNA fragmentation at the internucleosomal regions. Comparative study of the high molecular weight and internucleosomal DNA fragmentation demonstrated that these show different sensitivity to Zn2+ ions, actinomycin D and cycloheximide. In addition, the formation of large molecular weight fragments in contrast to oligonucleosomal ones may be reverted in conditions promoting to topoisomerase 11 mediated rejoining of cleaved DNA. These finding suggest that the high molecular weight and internucleosomal fragmentation may follow two different programВ¬mes of DNA cleavage during apoplosis mediated, presumably, by diverse enzymes. We showed that similar processes of ordered high molecular weight DNA cleavage occur in apoptotic cells and in those subjected to the stress challenges. The formation of high molecular weight DNA fragments was demonstrated to proceed promptly, show no correlation with cell death and occassionally may be of transient nature. The data obtained allow to interpret the formation of high molecular weight fragments as a component of cell response to stress rather that an early step of DNA fragmentation during apoptosis.
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References
[1]
Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251-306.
[2]
Arends MJ, Wyllie AH. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol. 1991;32:223-54.
[3]
Williams GT, Smith CA, McCarthy NJ, Grimes EA. Apoptosis: final control point in cell biology. Trends Cell Biol. 1992;2(9):263-7.
[4]
Cohen JJ, Duke RC, Fadok VA, Sellins KS. Apoptosis and programmed cell death in immunity. Annu Rev Immunol. 1992;10:267-93.
[5]
Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980;284(5756):555-6.
[6]
Compton MM. A biochemical hallmark of apoptosis: internucleosomal degradation of the genome. Cancer Metastasis Rev. 1992;11(2):105-19.
[7]
Walker PR, Kokileva L, LeBlanc J, Sikorska M. Detection of the initial stages of DNA fragmentation in apoptosis. Biotechniques. 1993;15(6):1032-40.
[8]
Brown DG, Sun XM, Cohen GM. Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. J Biol Chem. 1993;268(5):3037-9.
[9]
Oberhammer F, Wilson JW, Dive C, Morris ID, Hickman JA, Wakeling AE, Walker PR, Sikorska M. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 1993;12(9):3679-84.
[10]
Cohen GM, Sun XM, Fearnhead H, MacFarlane M, Brown DG, Snowden RT, Dinsdale D. Formation of large molecular weight fragments of DNA is a key committed step of apoptosis in thymocytes. J Immunol. 1994;153(2):507-16.
[11]
Heller C, Pohl FM. A systematic study of field inversion gel electrophoresis. Nucleic Acids Res. 1989;17(15):5989-6003.
[12]
Solov'ian VT, Andreev IO, Kunakh VA. Fractionation of eukaryotic DNA in a pulsed electrical field. II. Discrete DNA fragments and level of structural organization of chromatin. Mol Biol (Mosk). 1991;25(6):1483-91.
[13]
Nelson EM, Tewey KM, Liu LF. Mechanism of antitumor drug action: poisoning of mammalian DNA topoisomerase II on DNA by 4'-(9-acridinylamino)-methanesulfon-m-anisidide. Proc Natl Acad Sci U S A. 1984;81(5):1361-5.
[14]
Osheroff N. Biochemical basis for the interactions of type I and type II topoisomerases with DNA. Pharmacol Ther. 1989;41(1-2):223-41.
[16]
Kaufmann SH. Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: a cautionary note. Cancer Res. 1989;49(21):5870-8.
[17]
Ucker DS, Obermiller PS, Eckhart W, Apgar JR, Berger NA, Meyers J. Genome digestion is a dispensable consequence of physiological cell death mediated by cytotoxic T lymphocytes. Mol Cell Biol. 1992;12(7):3060-9.
[18]
Filipski J, Leblanc J, Youdale T, Sikorska M, Walker PR. Periodicity of DNA folding in higher order chromatin structures. EMBO J. 1990;9(4):1319-27.
[19]
Razin SV, Petrov P, Hancock R. Precise localization of the alpha-globin gene cluster within one of the 20- to 300-kilobase DNA fragments released by cleavage of chicken chromosomal DNA at topoisomerase II sites in vivo: evidence that the fragments are DNA loops or domains. Proc Natl Acad Sci U S A. 1991;88(19):8515-9.
[20]
Solovyan VT, Andreyev IO, Kunakh VA. The functional organization of plant nuclear DNA. I. Evidence for nuclear topoisomerase II. DNA complex. Mol Biol (Mosk). 1993; 27(6):1245-51.