Activation of the kinase activity of ATM by retinoic acid is required for CREB-dependent differentiation of neuroblastoma cells
Activation of the kinase activity of ATM by retinoic acid is required for CREB-dependent differentiation of neuroblastoma cells
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DOI：
10.1074/jbc.m609628200
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发表时间：
2007-06-01
影响因子：
4.8
通讯作者：
Price, Brendan D.
中科院分区：
文献类型：
--
作者：
Fernandes, Norvin D.;Sun, Yingli;Price, Brendan D.
ATM蛋白激酶在共济失调毛细血管扩张症中发生突变，共济失调毛细血管扩张症是一种以DNA修复缺陷、神经变性和生长因子信号传导缺陷为特征的遗传性疾病。ATM激酶的活性被DNA损伤激活，这种激活是细胞在遗传毒性事件中存活所必需的。除了在DNA修复中的这一良好特征作用外，我们现在证明了ATM在视黄酸（RA）诱导的SH-SY 5 Y神经母细胞瘤细胞分化为有丝分裂后的神经元样细胞中的新作用。RA快速激活ATM激酶的活性，导致CREB蛋白的ATM依赖性磷酸化，神经炎过程的挤出，以及SH-SY 5 Y细胞分化为神经元样细胞。当ATM蛋白表达被短发夹RNA抑制时，ATM依赖的CREB磷酸化被阻断。此外，当暴露于视黄酸时，ATM阴性细胞未能分化成神经元样细胞;相反，它们经历细胞死亡。组成型活性CREBVP 16构建体的表达，或暴露于毛喉素以诱导CREB磷酸化，挽救了ATM阴性细胞并恢复了分化。此外，当将CREB磷酸化位点（CREBS 133 A）或DNA结合结构域（KCREB）突变的显性负性CREB蛋白引入SH-SY 5 Y细胞时，维甲酸诱导的分化被阻断，细胞发生细胞死亡。结果表明，ATM是维甲酸诱导SH-SY 5 Y细胞分化所必需的，它通过依赖ATM的CREB丝氨酸133磷酸化而实现。因此，这些结果定义了一种新的机制，激活ATM激酶的活性，RA诱导分化过程中的CREB功能的调节，并牵连ATM。
The ATM protein kinase is mutated in ataxia telangiectasia, a genetic disease characterized by defective DNA repair, neurodegeneration, and growth factor signaling defects. The activity of ATM kinase is activated by DNA damage, and this activation is required for cells to survive genotoxic events. In addition to this well characterized role in DNA repair, we now demonstrate a novel role for ATM in the retinoic acid ( RA)induced differentiation of SH-SY5Y neuroblastoma cells into post-mitotic, neuronal-like cells. RA rapidly activates the activity of ATM kinase, leading to the ATM-dependent phosphorylation of the CREB protein, extrusion of neuritic processes, and differentiation of SH-SY5Y cells into neuronal-like cells. When ATM protein expression was suppressed by short hairpin RNA, the ATM-dependent phosphorylation of CREB was blocked. Furthermore, ATM-negative cells failed to differentiate into neuronal-like cells when exposed to retinoic acid; instead, they underwent cell death. Expression of a constitutively active CREBVP16 construct, or exposure to forskolin to induce CREB phosphorylation, rescued ATM negative cells and restored differentiation. Furthermore, when dominant negative CREB proteins with mutations in either the CREB phosphorylation site (CREBS133A) or the DNA binding domain (KCREB) were introduced into SH-SY5Y cells, retinoic acid-induced differentiation was blocked and the cells underwent cell death. The results demonstrate that ATM is required for the retinoic acid-induced differentiation of SH-SY5Y cells through the ATM dependent-phosphorylation of serine 133 of CREB. These results therefore define a novel mechanism for activation of the activity of ATM kinase by RA, and implicate ATM in the regulation of CREB function during RA-induced differentiation.