Previous studies have demonstrated that two Parkinson'sdisease-associatedgenes PINK1 and Parkin play a key role in mitochondrial quality control. But until now, the transcriptional regulation of these two genes under normal physiological conditions are not well understood. In this study, the transcriptional regulation of PINK1 and Parkin genes by nuclear respiratory factor 1 (NRF-1) and its effect on PINK1/ Parkin-mediated mitophagy were studied. The NRF-1 binding sites in the promoter regions of human PINK1 and Parkin genes were analyzed by JASPER software and were confirmed by chromatin immunoprecipitation (ChIP) assay. The transcriptional activities and the expressions of PINK1 and Parkin genes were positively regulated by NRF-1 in HEK293T cells and in SH-SY5Y cells. Furthermore, NRF-1 over-expression (OE) up-regulated the protein level of full-length PINK1 in CCCP-treated cells, indicating the enhanced PINK1/Parkin-mediated mitophagy. When NRF-1 expression was transient orstableknockdown, the CCCP-induced mitophagy was alleviated as characterized by the reduced protein level of full-length PINK1, the declined ratio of LC3 II to LC3 I, and the decreasedratioof Mt -keimafluorescenceintensityexcited at 552 nm to that excited at 488 nm. In conclusion, NRF-1 has a positive regulatory effect on the transcription of PINK1 and Parkin genes, and involves in mitochondrial quality control through regulating PINK1/Parkin-mediated mitophagy.

中文翻译：

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核呼吸因子1对人PINK1和Parkin基因表达的正调控

先前的研究表明，两个帕金森病相关基因 PINK1 和 Parkin 在线粒体质量控制中起着关键作用。但直到现在，这两个基因在正常生理条件下的转录调控还不是很清楚。本研究研究了核呼吸因子1（NRF-1）对PINK1和Parkin基因的转录调控及其对PINK1/Parkin介导的线粒体自噬的影响。人 PINK1 和 Parkin 基因启动子区域的 NRF-1 结合位点通过 JASPER 软件进行分析，并通过染色质免疫沉淀 (ChIP) 测定进行确认。NRF-1在HEK293T细胞和SH-SY5Y细胞中正调控PINK1和Parkin基因的转录活性和表达。此外，NRF-1 过表达 (OE) 上调 CCCP 处理细胞中全长 PINK1 的蛋白质水平，表明增强的 PINK1/Parkin 介导的线粒体自噬。当 NRF-1 表达被瞬时或稳定敲低时，CCCP 诱导的线粒体自噬得到缓解，其特征是全长 PINK1 的蛋白水平降低，LC3 II 与 LC3 I 的比率下降，以及在 552 nm 处激发的 Mt-keima 荧光强度与在 488 nm 激发。总之，NRF-1对PINK1和Parkin基因的转录具有正向调控作用，通过调控PINK1/Parkin介导的线粒体自噬参与线粒体质量控制。CCCP 诱导的线粒体自噬得到缓解，其特征是全长 PINK1 的蛋白质水平降低，LC3 II 与 LC3 I 的比率下降，以及在 552 nm 处激发的 Mt-keima 荧光强度与在 488 nm 处激发的强度的比率降低。总之，NRF-1对PINK1和Parkin基因的转录具有正向调控作用，通过调控PINK1/Parkin介导的线粒体自噬参与线粒体质量控制。CCCP 诱导的线粒体自噬得到缓解，其特征是全长 PINK1 的蛋白质水平降低，LC3 II 与 LC3 I 的比率下降，以及在 552 nm 处激发的 Mt-keima 荧光强度与在 488 nm 处激发的强度的比率降低。总之，NRF-1对PINK1和Parkin基因的转录具有正向调控作用，通过调控PINK1/Parkin介导的线粒体自噬参与线粒体质量控制。