Degradation of topoisomerase II alpha during adenovirus E1A-induced apoptosis is mediated by the activation of the ubiquitin proteolysis system

被引:51
作者
Nakajima, T
Morita, K
Ohi, N
Arai, T
Nozaki, N
Kikuchi, A
Osaka, F
Yamao, F
Oda, K
机构
[1] SCI UNIV TOKYO,DEPT APPL BIOL SCI,NODA,CHIBA 278,JAPAN
[2] KANAGAWA DENT COLL,DEPT ORAL BIOCHEM,YOKOSUKA,KANAGAWA 238,JAPAN
[3] MITSUBISHI CHEM INST LIFE SCI,MACHIDA,TOKYO 194,JAPAN
[4] NATL INST GENET,MISHIMA,SHIZUOKA 411,JAPAN
关键词
D O I
10.1074/jbc.271.40.24842
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The human epithermoid carcinoma-derived cell line MA1, established by introduction of the adenovirus E1A 12 S cDNA linked to the mouse mammary tumor virus long terminal repeat, elicits apoptosis after induction of E1A(12S) in response to dexamethasone. The level of topoisomerase II alpha begins to decrease steeply within 36 h preceding the onset of DNA fragmentation, whereas its mRNA level is unchanged (Nakajima, T., Ohi, N., Arai, T., Nozaki, N., Kikuchi, A., and Oda, K. (1995) Oncogene 10, 651-662). Topoisomerase II alpha prepared by immunoprecipitation or extraction of the nuclear matrix was degraded much more efficiently in the S10 extract prepared from MA1 cells treated with dexamethasone for 42 h (the 42-h extract) than in the extract from untreated MA1 cells (the 0-h extract) in an ATP- and ubiquitin-dependent manner. The proteolytic activity for degradation of topoisomerase II alpha was suppressed specifically by inhibitors for the proteasome and was much reduced in the 42-h extract prepared from MA1-derivative cell lines expressing E1B19k or Bcl-2. The proteolytic activity was lost after fractionation of the 42-h S10 extract into the S70 and P70 fractions by centrifugation at 70,000 x g for 6 h but partially recovered when these fractions were combined. Polyubiquitinated forms of topoisomerase II alpha could be detected by incubating it in the S70 or S100 extract, which lacks most of the proteasome activity. The ubiquitination activity in S70 prepared from the 42-h extract was 4- to 5-fold higher than that prepared from the 0-h extract. These results suggest that a component(s) in the ubiquitin proteolysis pathway, responsible for ubiquitination and degradation of topoisomerase II alpha, is activated or induced during the latent phase of E1A-induced apoptosis.
引用
收藏
页码:24842 / 24849
页数:8
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