PolyADP-ribosylation of histone H1 during apoptosis induced by DNA damaging agents in HL 60 cells = DNA 손상 인자에 의한 HL 60 세포의 예정사에서 histone H1 polyADP-ribosylation의 증가

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The biochemical role of polyADP-ribosylation on the internucleosomal DNA fragmentation associated with apoptosis was investigated in HL 60 human premyelocytic leukemia cells. An increase in the activity of poly(ADP-ribose)polymerase preceded DNA fragmentation during the UV-induced apoptotic process. The DNA fragmentation was prevented by inhibitors of poly(ADP-ribose)polymerase, suggesting a requirement of polyADP-ribosylation for DNA fragmentation. There was no significant increase of intracellular $Ca^{2+}$ level and $Ca^{2+}/Mg^{2+}$-dependent nuclease activity during UV-induced apoptosis. Intracellular level of $NAD^+$ did not changed significantly. It was found that UV light and chemotherapeutic drugs including adriamycin, mitomycin C and cisplatin increased polyADP-ribosylation of nuclear proteins, particularly histone H1. A poly(ADP-ribose)polymerase inhibitor, 3-aminobenzamide, prevented both internucleosomal DNA fragmentation and histone H1 polyADP-ribosylation in cells treated with these apoptosis inducers. These results suggest a correlation between the polyADP-ribosylation of histone H1 and internucleosomal DNA fragmentation in the apoptotic processes induced by DNA damaging agents. Our suggestion is further supported by the finding that a xeroderma pigmentosum cell line, GM04312B, which is defective in introducing nicks at the site of DNA damage, failed to induce DNA fragmentation as well as histone H1 polyADP-ribosylation after UV irradiation. To investigate the possible role of histone H1 polyADP-ribosylation, the micrococcal nuclease-susceptibility of chromatin was measured. Nuclease-susceptibility of chromatin was increased during UV-induced apoptosis, and the cotreatment of 3-aminobenzamide reduced both polyADP-ribosylation of histone H1 and nuclease-susceptibility of chromatin. A23187, colchicine and cycloheximide which do not induce DNA damage and PARP activation also provoked apoptosis in HL 60 cells. Apoptotic processes induced by these ag...
Joe, Cheol-Oresearcher조철오researcher
한국과학기술원 : 생물과학과,
Issue Date
105372/325007 / 000925232

학위논문(박사) - 한국과학기술원 : 생물과학과, 1996.2, [ x, 97 p. ]


DNA damage; polyADP-ribosylation; histone H1; Apoptosis; 핵산 손상; 에이디피리보실화; 히스톤; 예정사

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