Kwon Osong, Kim Kyung A, He Long, Kim Sun Ok, Kim Min Soo, Cha Eun Young, Yoon Byung Dae, Sok Dai Eun, Jung Mira, Ahn Jong Seog, Kim Bo Yeon
Korea Research Institute of Bioscience and Biotechnology, Yuseong, Daejeon, 305-806, Republic of Korea.
Cell Signal. 2008 Apr;20(4):602-12. doi: 10.1016/j.cellsig.2007.10.022. Epub 2007 Nov 6.
DNA damage by ionizing radiation (IR) can induce activations of both NF-kappaB and p53 through the upstream kinase ataxia telangiectasia mutated (ATM). NF-kappaB activation could also be signaled through two distinct or overlapped pathways; IkappaB kinases (IKKs)-IkappaBalpha and Akt-glycogen synthase kinase-3 (GSK-3). In the present study, however, we show that activation of Akt1 and the subsequent phosphorylation and inactivation of GSK-3beta by IR could also occur in ATM-deficient AT5BIVA cells as well as in normal MRC5CV1 fibroblasts. Similarly, lithium chloride (LiCl) was found to increase the phosphorylation of GSK-3beta independently of ATM. Transfection with either wild-type or kinase dead mutant GSK-3beta to the cells further indicated that phosphorylations of Akt1 and GSK-3beta were closely associated with the transcriptional transactivation of NF-kappaB in response to ionizing radiation. On the other hand, LiCl, having no effect on caspase-3 activation, significantly increased p53 phosphorylation and apoptotic death of the normal MRC5CV1 cells while IR, activating both caspase-3 and p53, profoundly affected AT5BIVA cell death. Hence, our data suggest that although ATM-mediated IKK-IkappaBalpha pathway might be a typical pathway for IR-induced NF-kappaB activation, GSK-3beta phosphorylation could also partially contribute to the transcriptional transactivation of NF-kappaB in an ATM-independent manner and that GSK-3beta phosphorylation could induce ATM-mediated cell apoptosis through the activation of p53.
电离辐射(IR)造成的DNA损伤可通过上游激酶共济失调毛细血管扩张症突变基因(ATM)诱导核因子-κB(NF-κB)和p53激活。NF-κB激活也可通过两条不同或重叠的途径发出信号;IκB激酶(IKK)-IκBα和Akt-糖原合酶激酶-3(GSK-3)。然而,在本研究中,我们发现IR诱导的Akt1激活以及随后GSK-3β的磷酸化和失活,在ATM缺陷的AT5BIVA细胞以及正常的MRC5CV1成纤维细胞中也会发生。同样,发现氯化锂(LiCl)可独立于ATM增加GSK-3β的磷酸化。用野生型或激酶失活突变体GSK-3β转染细胞进一步表明,Akt1和GSK-3β的磷酸化与电离辐射后NF-κB的转录反式激活密切相关。另一方面,LiCl对半胱天冬酶-3激活没有影响,但显著增加了正常MRC5CV1细胞的p53磷酸化和凋亡死亡,而IR同时激活半胱天冬酶-3和p53,对AT5BIVA细胞死亡有深远影响。因此,我们的数据表明,尽管ATM介导的IKK-IκBα途径可能是IR诱导NF-κB激活的典型途径,但GSK-3β磷酸化也可能以不依赖ATM的方式部分促进NF-κB的转录反式激活,并且GSK-3β磷酸化可通过激活p53诱导ATM介导的细胞凋亡。