Genome Research, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto-ku, Tokyo 135-8550, Japan.
Genome Research, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto-ku, Tokyo 135-8550, Japan
J Biol Chem. 2020 Apr 3;295(14):4591-4603. doi: 10.1074/jbc.RA119.010655. Epub 2020 Feb 27.
As a branch of the unfolded protein response, protein kinase R-like endoplasmic reticulum kinase (PERK) represses global translation in response to endoplasmic reticulum (ER) stress. This pathophysiological condition is associated with the tumor microenvironment in cancer. Previous findings in our lab have suggested that PERK selectively represses translation of some mRNAs, but this possibility awaits additional investigation. In this study, we show that a stem-cell marker protein, leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), is rapidly depleted in colon cancer cells during ER stress, an effect that depended on the PERK-mediated translational repression. Indeed, the PERK inhibition led to the accumulation of premature, underglycosylated forms of LGR5, which were produced only at low levels during proper PERK activation. Unlike the mature LGR5 form, which is constitutively degraded regardless of PERK activation, the underglycosylated LGR5 exhibited a prolonged half-life and accumulated inside the cells without being expressed on the cell surface. We also found that Erb-B2 receptor tyrosine kinase 3 (ERBB3) is subjected to a similarly-regulated depletion by PERK, whereas the epidermal growth factor receptor (EGFR), stress-inducible heat-shock protein family A (Hsp70) member 5 (HSPA5), and anterior gradient 2 protein-disulfide isomerase family member (AGR2) were relatively. insensitive to the PERK-mediated repression of translation. These results indicate that LGR5 and ERBB3 are targets for PERK-mediated translational repression during ER stress.
作为未折叠蛋白反应的一个分支,蛋白激酶 R 样内质网激酶 (PERK) 响应内质网 (ER) 应激抑制翻译的全局水平。这种病理生理状况与肿瘤微环境中的癌症有关。我们实验室之前的研究结果表明,PERK 选择性地抑制一些 mRNA 的翻译,但这一可能性有待进一步研究。在这项研究中,我们发现干细胞标志物蛋白富含亮氨酸重复的 G 蛋白偶联受体 5(LGR5)在 ER 应激期间迅速耗尽结肠癌细胞,这种效应依赖于 PERK 介导的翻译抑制。事实上,PERK 抑制导致 LGR5 的过早、低聚糖形式的积累,而在适当的 PERK 激活期间,仅以低水平产生。与成熟的 LGR5 形式不同,后者无论 PERK 激活如何都被持续降解,而过糖基化的 LGR5 形式半衰期延长,并在细胞内积累而不表达在细胞表面。我们还发现,表皮生长因子受体酪氨酸激酶 3 (ERBB3) 也受到 PERK 类似调节的消耗,而表皮生长因子受体 (EGFR)、应激诱导的热休克蛋白家族 A (Hsp70) 成员 5 (HSPA5) 和前梯度 2 蛋白二硫键异构酶家族成员 (AGR2) 则相对不受 PERK 介导的翻译抑制的影响。这些结果表明,LGR5 和 ERBB3 是 PERK 介导的 ER 应激期间翻译抑制的靶标。