Abstract
Interaction of bleomycin with nuclei isolated from a variety of mammalian cells resulted in the release of nucleosomes. When isolated mononucleosomes (core plus linker) were re-treated with bleomycin, no further degradation of DNA occurred. The results suggest that the bleomycin cleavage sites in chromatin are present only in the linker region and that there are probably only one or two cleavage sites per linker. The repeat lengths of nucleosomal DNA released by bleomycin from nuclei of different species are different; this variability is considered to reflect the length of the linker. Incorporation of BrdU into DNA did not alter the bleomycin action on nucleosomes. When mitotic cells were held at metaphase for a prolonged period, bleomycin caused a gradual disintegration of chromosomes, although the bleomycin cleavage sites in metaphase chromosomes were found to be the same as those in interphase nuclei.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akiyama, Y., Kato, S.: Two cell lines from lymphomas of Marek's disease. Biken J. 17, 105–116 (1974)
Blum, R.H., Carter, S.K., Agre, K.: A clinical review of bleomycin—a new antineoplastic agent. Cancer (Philad.) 31, 903–914 (1973)
Brack, C, Eberle, H., Bickle, T.A., Yuan, R.: A map of the sites of bacteriophage PM2 DNA for the restriction endonucleases HindIII and HpaII. J. molec. Biol. 104, 305–309 (1976)
Chambon, P.: Molecular biology of eukaryotic genome is coming of age. Cold Spr. Harb. Symp. quant. Biol. (in press, 1977)
Compton, J.L., Bellard, M., Chambon. P.: Biochemical evidence of variability in the DNA repeated length in the chromatin of higher eukaryotes. Proc. nat. Acad. Sci. (Wash.) 73, 4382–4386 (1976a)
Compton, J.L., Hancock, R., Oudet, P., Chambon, P.: Biochemical and electron microscopic evidence that the subunit structure of Chinese hamster ovary interphase chromatin is conserved in mitotic chromosomes. Europ. J. Biochem. 70, 555–568 (1976b)
Felsenfeld, G.: Chromatin. Nature (Lond.) 271, 115–122 (1978)
Finch, J.T., Lutter, L.C., Rhodes, D., Brown, R.S., Rushton, B., Levitt, M., Klug, A.: Structure of nucleosome core particles of chromatin. Nature (Lond.) 269, 29–36 (1977)
Haidle, C.W., Kuo, M.T., Weiss, K.K.: Nucleic acid-specificity of bleomycin. Biochem. Pharmacology 21, 3308–3312 (1972b)
Haidle, C.W., Weiss, K.K., Kuo, M.T.: Release of free bases from deoxyribonucleic acid after reaction with bleomycin. Molec. Pharmacology 8, 531–537 (1972a)
Hittleman, W.M., Rao, P.N.: Bleomycin-induced damage in prematurely condensed chromosomes and its relationship to cell cycle progression in CHO cells. Cancer Res. 34, 3433–3439 (1974)
Ishida, R., Takahashi, T.: Increased DNA strand breakage by combined action of bleomycin and superoxide radical. Biochem. biophys. Res. Commun. 66, 1432–1438 (1975)
Kornberg, R.A.: Structure of chromatin. Ann. Rev. Biochem. 46, 931–954 (1977)
Kovacic, R.T., Holde, K.E. Van: Sedimentation of homogenous double strand DNA molecules. Biochemistry 16, 1490–1498 (1977)
Kuo, M.T., Auger, L.A., Saunders, G.F., Haidle, C.W.: Effect of bleomycin on the synthesis and function of DNA. Cancer Res. 37, 1345–1348 (1977)
Kuo, M.T., Haidle, C.W.: Characterization of chain breakage in DNA induced by bleomycin. Biochem. biophys. Acta (Amst.) 335, 109–114 (1973)
Kuo, M.T., Hsu, T.C.: Bleomycin causes release of nucleosomes from chromatin and chromosomes. Nature (Lond.) 271, 83–84 (1978)
Kuo, M.T., Sahasrabuddhe, C.G., Saunders, G.F.: Presence of messenger specifying sequences in the DNA of chromatin subunits. Proc. nat. Acad. Sci. (Wash.) 73, 1572–1575 (1976)
Lloyd, R.S., Haidle, C.W., Robberson, D.L.: Bleomycin-specific fragmentation of double-stranded DNA. Biochemistry (in press, 1978)
Luk, D.C., Bick, M.D.: Determination of 5′-bromodeoxyuridine in DNA by buoyant density. Anal. Biochem. 77, 346–349 (1977)
Maniatis, T., Jefferey, A., van de Sande, H.: Chain length determination of small double and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14, 3787–3794 (1975)
McDonell, M.W., Simon, M.N., Studier, F.W.: Analysis of restriction fragments of T7 DNA and Determination of Molecular Weights by electrophoresis in neutral and alkaline gels. J. molec. Biol. 110, 119–146 (1977)
Müller, W.E.G., Yamazaki, Z., Breter, H.J., Zahn, R.K.: Action of bleomycin on DNA and RNA. Europ. J. Biochem. 31, 518–525 (1972)
Müller, W.E.G., Zahn, R.K.: Bleomycin, an antibiotic that removes thymine from double-stranded DNA. Prog. Nucl. Acid. Res. molec. Biol. 40, 21–57 (1977)
Ohama, K., Kadotani, T.: Cytologic effects of bleomycin on cultured human leucocytes. Jap. J. Hum. Genet. 14, 293–297 (1970)
Paika, K.D., Krishan, A.: Bleomycin-induced chromosomal aberrations in cultured mammalian cells. Cancer Res. 33, 961–965 (1973)
Parker, R.C., Watson, R.M., Vinograd, J.: Mapping of closed circular DNA by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis. Proc. nat. Acad. Sci. (Wash.) 74, 851–855 (1977)
Suzuki, H., Nagai, K., Akutsu, E., Yamaki, H., Tanaka, N., Umezawa, H.: On the mechanism of action of bleomycin, strand scission of DNA caused by bleomycin and its binding to DNA in vitro. J. Antibiot. (Tokyo) 23, 473–480 (1970)
Umezawa, H., Asakura, H., Oda, K., Hori, S., Hori, M.: The effect of bleomycin on SV40 DNA: characteristics of bleomycin action which produces a single scission in a superhelical form of SV40 DNA. J. Antibiot. (Tokyo) 26, 521–527 (1973)
Umezawa, H., Maeda, K., Takeuchi, T., Okami, Y.: New Antibiotics, bleomycin A and B. J. Antibiot. (Tokyo) 19, 200–209 (1966)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kuo, M.T., Hsu, T.C. Biochemical and cytological studies of bleomycin actions on chromatin and chromosomes. Chromosoma 68, 229–240 (1978). https://doi.org/10.1007/BF00335418
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00335418