Positive regulation of human PINK1 and Parkin gene expression by nuclear respiratory factor 1
Positive regulation of human PINK1 and Parkin gene expression by nuclear respiratory factor 1
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核呼吸因子1对人类PINK1和Parkin基因表达的正向调节
DOI：
10.1016/j.mito.2019.12.002
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发表时间：
2020-03-01
期刊：
影响因子：
4.4
通讯作者：
Zhu, Li
中科院分区：
文献类型：
--
作者：
Lu, Yapeng;Ding, Wangwang;Zhu, Li
先前的研究表明，两个帕金森病相关基因PINK 1和Parkin在线粒体质量控制中起着关键作用。但到目前为止，这两个基因在正常生理条件下的转录调控还不是很清楚。本研究探讨了核呼吸因子1（NRF-1）对PINK 1和Parkin基因的转录调控及其对PINK 1/Parkin介导的线粒体自噬的影响。用碧玉软件分析人PINK 1和Parkin基因启动子区的NRF-1结合位点，并通过染色质免疫沉淀（ChIP）试验进行验证。在HEK 293 T细胞和SH-SY 5 Y细胞中，NRF-1对PINK 1和Parkin基因的转录活性和表达具有正调控作用。此外，NRF-1过表达（OE）上调CCCP处理的细胞中全长PINK 1的蛋白水平，表明PINK 1/Parkin介导的线粒体自噬增强。当NRF-1表达被瞬时或稳定敲低时，CCCP诱导的线粒体自噬现象减轻，表现为全长PINK 1蛋白水平降低，LC 3 II/LC 3 I比值降低，552 nm激发的Mtkeima荧光强度与488 nm激发的荧光强度比值降低。总之，NRF-1对PINK 1和Parkin基因的转录具有正调控作用，并通过调节PINK 1/Parkin介导的线粒体自噬参与线粒体质量控制。
Previous studies have demonstrated that two Parkinson's disease-associated genes 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 or stable knockdown, 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 decreased ratio of Mtkeima fluorescence intensity excited 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.