Department of Public Health, School of Pharmacy, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.
Sci Rep. 2017 Dec 1;7(1):16735. doi: 10.1038/s41598-017-17112-8.
Methylmercury (MeHg) is a widely distributed environmental pollutant that causes a series of cytotoxic effects. However, molecular mechanisms underlying MeHg toxicity are not fully understood. Here, we report that sequestosome1/p62 protects mouse embryonic fibroblasts (MEFs) against low-dose MeHg cytotoxicity via clearance of MeHg-induced ubiquitinated proteins. p62 mRNA and protein expression in MEFs were temporally induced by MeHg exposure p62-deficient MEFs exhibited higher sensitivity to MeHg exposure compared to their wild-type (WT) counterparts. An earlier and higher level of accumulation of ubiquitinated proteins was detected in p62-deficient cells compared with WT MEFs. Confocal microscopy revealed that p62 and ubiquitinated proteins co-localized in the perinuclear region of MEFs following MeHg treatment. Further analysis of MEFs revealed that ubiquitinated proteins co-localized with LC3-positive puncta upon co-treatment with MeHg and chloroquine, an autophagy inhibitor. In contrast, there was minimal co-localization in p62-deficient MEFs. The present study, for the first time, examined the expression and distribution of p62 and ubiquitinated proteins in cells exposed to low-dose MeHg. Our findings suggest that p62 is crucial for cytoprotection against MeHg-induced toxicity and is required for MeHg-induced ubiquitinated protein clearance.
甲基汞(MeHg)是一种广泛分布的环境污染物,可引起一系列细胞毒性作用。然而,甲基汞毒性的分子机制尚未完全阐明。在这里,我们报告了自噬相关蛋白 p62 通过清除 MeHg 诱导的泛素化蛋白来保护小鼠胚胎成纤维细胞(MEFs)免受低剂量 MeHg 的细胞毒性。MeHg 暴露可使 MEFs 中 p62 的 mRNA 和蛋白表达呈时间依赖性诱导。与野生型(WT)细胞相比,p62 缺陷型 MEFs 对 MeHg 暴露更为敏感。与 WT MEFs 相比,p62 缺陷型细胞中检测到的泛素化蛋白的积累更早且水平更高。共聚焦显微镜显示,MeHg 处理后 p62 和泛素化蛋白在 MEFs 的核周区域共定位。对 MEFs 的进一步分析表明,在用 MeHg 和氯喹(一种自噬抑制剂)共同处理时,泛素化蛋白与 LC3 阳性斑点共定位。相比之下,p62 缺陷型 MEFs 中几乎没有共定位。本研究首次研究了暴露于低剂量 MeHg 的细胞中 p62 和泛素化蛋白的表达和分布。我们的研究结果表明,p62 对于防止 MeHg 诱导的毒性具有重要作用,并且对于 MeHg 诱导的泛素化蛋白清除是必需的。