Howard Hughes Medical Institute, Robin Chemers Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States.
Institute for Regenerative Medicine, University of Zurich, Zurich, Switzerland.
Elife. 2022 Jun 27;11:e77780. doi: 10.7554/eLife.77780.
Cells encountering stressful situations activate the integrated stress response (ISR) pathway to limit protein synthesis and redirect translation to better cope. The ISR has also been implicated in cancers, but redundancies in the stress-sensing kinases that trigger the ISR have posed hurdles to dissecting physiological relevance. To overcome this challenge, we targeted the regulatory node of these kinases, namely, the S51 phosphorylation site of eukaryotic translation initiation factor eIF2α and genetically replaced eIF2α with eIF2α-S51A in mouse squamous cell carcinoma (SCC) stem cells of skin. While inconsequential under normal growth conditions, the vulnerability of this ISR-null state was unveiled when SCC stem cells experienced proteotoxic stress. Seeking mechanistic insights into the protective roles of the ISR, we combined ribosome profiling and functional approaches to identify and probe the functional importance of translational differences between ISR-competent and ISR-null SCC stem cells when exposed to proteotoxic stress. In doing so, we learned that the ISR redirects translation to centrosomal proteins that orchestrate the microtubule dynamics needed to efficiently concentrate unfolded proteins at the microtubule-organizing center so that they can be cleared by the perinuclear degradation machinery. Thus, rather than merely maintaining survival during proteotoxic stress, the ISR also functions in promoting cellular recovery once the stress has subsided. Remarkably, this molecular program is unique to transformed skin stem cells, hence exposing a vulnerability in cancer that could be exploited therapeutically.
细胞在遇到应激情况时会激活整合应激反应(ISR)途径,以限制蛋白质合成并重新定向翻译,以更好地应对。ISR 也与癌症有关,但触发 ISR 的应激感应激酶的冗余性给解析生理相关性带来了障碍。为了克服这一挑战,我们针对这些激酶的调节节点,即真核翻译起始因子 eIF2α 的 S51 磷酸化位点,并用 eIF2α-S51A 基因替代了皮肤鳞状细胞癌(SCC)干细胞中的 eIF2α。在正常生长条件下,这种 ISR 缺失状态没有影响,但当 SCC 干细胞受到蛋白毒性应激时,这种状态的脆弱性就暴露出来了。为了深入了解 ISR 的保护作用的机制,我们结合核糖体图谱和功能方法,在暴露于蛋白毒性应激时,识别并探测 ISR 功能完整和缺失的 SCC 干细胞之间翻译差异的功能重要性。通过这样做,我们了解到 ISR 将翻译重新定向到中心体蛋白,这些蛋白协调微管动力学,以便有效地将未折叠的蛋白质集中在微管组织中心,从而可以通过核周降解机制进行清除。因此,ISR 不仅在蛋白毒性应激期间维持存活,而且在应激消退后还能促进细胞恢复。值得注意的是,这种分子程序是转化的皮肤干细胞所特有的,因此揭示了癌症的一个弱点,可以在治疗上加以利用。