Lipskaia L A
Tsitologiia. 1994;36(3):303-9.
With the aim of revealing endonuclease(s), activating in apoptotic cells, a nuclease activity of nuclear protein extract from human leukemic HL-60 cells treated with colchicine was studied. After a 30 h incubation with colchicine, a specific DNA fragmentation--a multiple of 200 base pair subunits--was clearly observed that was an evidence for the endonuclease activation. After 48 h almost the whole DNA was fragmented. At the same time histones appeared the major proteins of nuclear extract. Using the method for detection of nuclease activity in a gel, protein 37 kDa was identified as a Ca2+, Mg(2+)-dependent endonuclease. The maximum increase in nuclease activity occurred after a 24 h incubation with colchicine. A low activity of this protein was also found in control cells. These data suggest that in apoptotic cells activation of constantly expressed nucleases may occur, rather than a new synthesis of apoptotic nucleases.
为了揭示在凋亡细胞中激活的核酸内切酶,对用秋水仙碱处理的人白血病HL-60细胞核蛋白提取物的核酸酶活性进行了研究。用秋水仙碱孵育30小时后,明显观察到一种特定的DNA片段化现象——由200个碱基对亚基组成的倍数——这是核酸内切酶激活的证据。48小时后,几乎所有的DNA都被片段化。与此同时,组蛋白成为核提取物中的主要蛋白质。使用凝胶中核酸酶活性检测方法,鉴定出37 kDa的蛋白质为一种依赖Ca2+、Mg2+的核酸内切酶。与秋水仙碱孵育24小时后,核酸酶活性达到最大增加。在对照细胞中也发现了这种蛋白质的低活性。这些数据表明,在凋亡细胞中可能发生持续表达的核酸酶的激活,而不是凋亡核酸酶的新合成。