Wright S C, Schellenberger U, Ji L, Wang H, Larrick J W
The Palo Alto Institute of Molecular Medicine, Mountain View, California 94043, USA.
FASEB J. 1997 Sep;11(11):843-9. doi: 10.1096/fasebj.11.11.9285482.
The present studies describe a new function for calmodulin-dependent protein kinase II (CaM-KII) in signal transduction leading to apoptosis. Both tumor necrosis factor alpha (TNF) and UV light rapidly stimulated Ca2+-independent activity of CaM-KII in the monocytic leukemia, U937. Two mechanistically different inhibitors of CaM-KII blocked activation of CaM-KII and prevented DNA fragmentation and death. Activation of CaM-KII during apoptosis and inhibition of DNA fragmentation by the two CaM-KII inhibitors were reproduced in several other lines including KGla, HL-60, and YAC-1. However, K562, which is relatively resistant to apoptosis induced by either TNF or UV light, did not activate CaM-KII in response to these stimuli. A variant derived from U937 that is resistant to TNF- or UV light-induced apoptosis also lacked a CaM-KII response. Activation of Cam-KII was blocked by two protease inhibitors, VAD-fmk and TPCK, but not by other inhibitors of serine proteases. Both inhibitors of CaM-KII and the protease inhibitors blocked activation of AP24, a serine protease originally isolated from apoptotic cells that induces DNA fragmentation in nuclei. Our evidence supports a model in which proteolytic activity functions upstream of CaM-KII. This kinase then leads to activation of AP24, which transmits signals to the nucleus to initiate DNA fragmentation.
目前的研究描述了钙调蛋白依赖性蛋白激酶II(CaM-KII)在导致细胞凋亡的信号转导中的新功能。肿瘤坏死因子α(TNF)和紫外线均可快速刺激单核细胞白血病U937细胞中CaM-KII的非钙依赖性活性。两种机制不同的CaM-KII抑制剂可阻断CaM-KII的激活,并防止DNA片段化和细胞死亡。在包括KGla、HL-60和YAC-1在内的其他几种细胞系中也再现了细胞凋亡过程中CaM-KII的激活以及两种CaM-KII抑制剂对DNA片段化的抑制作用。然而,对TNF或紫外线诱导的细胞凋亡相对抗性的K562细胞,在受到这些刺激时不会激活CaM-KII。从U937衍生而来的对TNF或紫外线诱导的细胞凋亡具有抗性的变体也缺乏CaM-KII反应。CaM-KII的激活被两种蛋白酶抑制剂VAD-fmk和TPCK阻断,但未被其他丝氨酸蛋白酶抑制剂阻断。CaM-KII抑制剂和蛋白酶抑制剂均阻断了AP24的激活,AP24是一种最初从凋亡细胞中分离出来的丝氨酸蛋白酶,可诱导细胞核中的DNA片段化。我们的证据支持一种模型,即蛋白水解活性在CaM-KII的上游起作用。然后这种激酶导致AP24的激活,AP24将信号传递到细胞核以启动DNA片段化。