Kieran Dairín, Woods Ina, Villunger Andreas, Strasser Andreas, Prehn Jochen H M
Department of Physiology and Medical Physics and Royal College of Surgeons in Ireland Neuroscience Research Centre, Royal College of Surgeons in Ireland, 123 St. Stephen's Green, Dublin 2, Ireland.
Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20606-11. doi: 10.1073/pnas.0707906105. Epub 2007 Dec 11.
BH3-only proteins couple diverse stress signals to the evolutionarily conserved mitochondrial apoptosis pathway. Previously, we reported that the activation of the BH3-only protein p53-up-regulated mediator of apoptosis (Puma) was necessary and sufficient for endoplasmic reticulum (ER) stress- and proteasome inhibition-induced apoptosis in neuroblastoma and other cancer cells. Defects in protein quality control have also been suggested to be a key event in ALS, a fatal neurodegenerative condition characterized by motoneuron degeneration. Using the SOD1(G93A) mouse model as well as human post mortem samples from ALS patients, we show evidence for increased ER stress and defects in protein degradation in motoneurons during disease progression. Before symptom onset, we detected a significant up-regulation of Puma in motoneurons of SOD1(G93A) mice. Genetic deletion of puma significantly improved motoneuron survival and delayed disease onset and motor dysfunction in SOD1(G93A) mice. However, it had no significant effect on lifespan, suggesting that other ER stress-related cell-death proteins or other factors, such as excitotoxicity, necrosis, or inflammatory injury, may contribute at later disease stages. Indeed, further experiments using cultured motoneurons revealed that genetic deletion of puma protected motoneurons against ER stress-induced apoptosis but showed no effect against excitotoxic injury. These findings demonstrate that a single BH3-only protein, the ER stress-associated protein Puma, plays an important role during the early stages of chronic neurodegeneration in vivo.
仅含BH3结构域的蛋白将多种应激信号与进化上保守的线粒体凋亡途径相偶联。此前,我们报道过,仅含BH3结构域的蛋白p53上调凋亡调节因子(Puma)的激活对于内质网(ER)应激和蛋白酶体抑制诱导的神经母细胞瘤及其他癌细胞凋亡而言是必要且充分的。蛋白质质量控制缺陷也被认为是肌萎缩侧索硬化症(ALS)中的一个关键事件,ALS是一种以运动神经元变性为特征的致命神经退行性疾病。利用SOD1(G93A)小鼠模型以及ALS患者的人类尸检样本,我们发现了疾病进展过程中运动神经元内质网应激增加和蛋白质降解缺陷的证据。在症状出现之前,我们检测到SOD1(G93A)小鼠运动神经元中Puma显著上调。puma基因缺失显著提高了运动神经元的存活率,并延迟了SOD1(G93A)小鼠的疾病发作和运动功能障碍。然而,它对寿命没有显著影响,这表明其他内质网应激相关的细胞死亡蛋白或其他因素,如兴奋性毒性、坏死或炎性损伤,可能在疾病后期起作用。事实上,使用培养的运动神经元进行的进一步实验表明,puma基因缺失可保护运动神经元免受内质网应激诱导的凋亡,但对兴奋性毒性损伤没有影响。这些发现表明,单一的仅含BH3结构域的蛋白,即内质网应激相关蛋白Puma,在体内慢性神经退行性变的早期阶段发挥重要作用。