Jack Melissa T, Woo Richard A, Hirao Atsushi, Cheung Alison, Mak Tak W, Lee Patrick W K
Cancer Biology Research Group and Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Center, Calgary, AL, Canada T2N 4N1.
Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9825-9. doi: 10.1073/pnas.152053599. Epub 2002 Jul 3.
In response to genotoxic stress, mammalian cells can activate cell cycle checkpoint pathways to arrest the cell for repair of DNA damage or induce apoptosis to eliminate damaged cells. The checkpoint kinase, Chk2, has been implicated in both of these responses and is believed to function in an ataxia telangiectasia (Atm)-dependent manner. We show here that Chk2-/- mouse embryo fibroblasts (MEFs), unlike Atm-/- or p53-/- MEFs, behaved like normal MEFs in manifesting p21 induction and G(1) arrest upon exposure to gamma-irradiation. Therefore, Chk2 is not involved in p53-mediated G(1) arrest. To examine the role of Chk2 in p53-dependent apoptotic response, we used adenovirus E1A-expressing MEFs. We show that Chk2-/- cells, like p53-/- cells, did not undergo DNA damage-induced apoptosis, whereas Atm-/- cells behaved like normal cells in invoking an apoptotic response. Furthermore, this apoptosis could occur in the absence of protein synthesis, suggesting that it is preexisting, or "latent," p53 that mediates this response. We conclude that Chk2 is not involved in Atm- and p53-dependent G(1) arrest, but is involved in the activation of latent p53, independently of Atm, in triggering DNA damage-induced apoptosis.
为应对基因毒性应激,哺乳动物细胞可激活细胞周期检查点通路,使细胞停滞以修复DNA损伤,或诱导细胞凋亡以清除受损细胞。检查点激酶Chk2与这两种反应均有关联,且被认为以共济失调毛细血管扩张症(Atm)依赖的方式发挥作用。我们在此表明,与Atm-/-或p53-/-小鼠胚胎成纤维细胞(MEF)不同,Chk2-/- MEF在受到γ射线照射后表现出p21诱导和G(1)期停滞,其行为与正常MEF相似。因此,Chk2不参与p53介导的G(1)期停滞。为研究Chk2在p53依赖的凋亡反应中的作用,我们使用了表达腺病毒E1A的MEF。我们发现,Chk2-/-细胞与p53-/-细胞一样,不会发生DNA损伤诱导的凋亡,而Atm-/-细胞在引发凋亡反应时表现得与正常细胞相似。此外,这种凋亡可在无蛋白质合成的情况下发生,这表明介导此反应的是预先存在的或“潜伏”的p53。我们得出结论,Chk2不参与Atm和p53依赖的G(1)期停滞,但在触发DNA损伤诱导的凋亡过程中,独立于Atm参与潜伏p53的激活。