Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Japan.
FEBS Open Bio. 2023 Sep;13(9):1667-1682. doi: 10.1002/2211-5463.13682. Epub 2023 Aug 8.
Telomerase reverse transcriptase (TERT) is a protein that catalyzes the reverse transcription of telomere elongation. TERT is also expected to play a non-canonical role beyond telomere lengthening since it localizes not only in the nucleus but also in mitochondria, where telomeres do not exist. Several studies have reported that mitochondrial TERT regulates apoptosis induced by oxidative stress. However, there is still some controversy as to whether mitochondrial TERT promotes or inhibits apoptosis, mainly due to the lack of information on changes in TERT distribution in individual cells over time. Here, we simultaneously detected apoptosis and TERT localization after oxidative stress in individual HeLa cells by live-cell tracking. Single-cell tracking revealed that the stress-induced accumulation of TERT in mitochondria caused apoptosis, but that accumulation increased over time until cell death. The results suggest a new model in which mitochondrial TERT has two opposing effects at different stages of apoptosis: it predetermines apoptosis at the first stage of cell-fate determination, but also delays apoptosis at the second stage. As such, our data support a model that integrates the two opposing hypotheses on mitochondrial TERT's effect on apoptosis. Furthermore, detailed statistical analysis of TERT mutations, which have been predicted to inhibit TERT transport to mitochondria, revealed that these mutations suppress apoptosis independent of mitochondrial localization of TERT. Together, these results imply that the non-canonical functions of TERT affect a wide range of mitochondria-dependent and mitochondria-independent apoptosis pathways.
端粒酶逆转录酶 (TERT) 是一种催化端粒延长的逆转录的蛋白质。由于 TERT 不仅定位于细胞核,还定位于不存在端粒的线粒体中,因此预计它将发挥非规范的作用,超越端粒延长。有几项研究报道,线粒体 TERT 调节氧化应激诱导的细胞凋亡。然而,由于缺乏关于 TERT 在单个细胞中随时间分布变化的信息,线粒体 TERT 是促进还是抑制细胞凋亡仍存在一些争议。在这里,我们通过活细胞追踪同时检测了个体 HeLa 细胞在氧化应激后细胞凋亡和 TERT 定位。单细胞追踪显示,应激诱导的 TERT 在线粒体中的积累导致细胞凋亡,但随着时间的推移,积累量增加,直至细胞死亡。结果表明,线粒体 TERT 在细胞命运决定的第一阶段预先确定细胞凋亡,但在第二阶段又延迟细胞凋亡,存在两种相反的作用。因此,我们的数据支持了一种整合了关于线粒体 TERT 对细胞凋亡影响的两种相反假说的模型。此外,对预测抑制 TERT 向线粒体运输的 TERT 突变的详细统计分析表明,这些突变独立于 TERT 的线粒体定位抑制细胞凋亡。综上所述,这些结果表明 TERT 的非规范功能影响广泛的线粒体依赖性和非线粒体依赖性细胞凋亡途径。