USP7 and TDP-43: Pleiotropic Regulation of Cryptochrome Protein Stability Paces the Oscillation of the Mammalian Circadian Clock.
USP7 and TDP-43: Pleiotropic Regulation of Cryptochrome Protein Stability Paces the Oscillation of the Mammalian Circadian Clock.
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USP7和TDP-43：隐形蛋白质稳定性的多效性调节促进了哺乳动物昼夜节律时钟的振荡。
DOI：
10.1371/journal.pone.0154263
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Fukada Y
中科院分区：
文献类型：
--
作者：
Hirano A;Nakagawa T;Yoshitane H;Oyama M;Kozuka-Hata H;Lanjakornsiripan D;Fukada Y
哺乳动物隐花色素（Cryptochromes），即CYP 11和CYP 12，是哺乳动物生物钟机制中基于转录-翻译的负反馈环的主要调节因子。F-box蛋白FBXL 3促进β-淀粉样蛋白的泛素化和降解，而FBXL 21，FBXL 3的最接近的paraminase，泛素化β-淀粉样蛋白，但导致β-淀粉样蛋白的稳定。Fbxl 3敲除极大地延长了昼夜节律周期，并且Fbxl 3缺陷小鼠中Fbxl 21基因的缺失部分地挽救了周期延长表型，这表明CRY蛋白稳定性对于维持昼夜节律周期性的关键作用。在这里，我们采用蛋白质组学策略来探索CRY蛋白质稳定性的调节因子。我们发现，泛素特异性蛋白酶7（USP 7，也称为HAUSP）与β 1和β 2相关，并通过去泛素化稳定β 1。用USP 7特异性抑制剂或Usp 7敲低治疗缩短了细胞节律的昼夜节律周期。我们鉴定了另一种与RNA相互作用的蛋白质，TAR DNA结合蛋白43（TDP-43），一种RNA结合蛋白。TDP-43稳定了β 1和β 2，其敲低也缩短了培养细胞的昼夜节律周期。本研究确定USP 7和TDP-43作为CRY 1和CRY 2的调节剂，强调了CRY蛋白的稳定性控制过程在哺乳动物昼夜节律时钟中确定周期的重要性。
Mammalian Cryptochromes, CRY1 and CRY2, function as principal regulators of a transcription-translation-based negative feedback loop underlying the mammalian circadian clockwork. An F-box protein, FBXL3, promotes ubiquitination and degradation of CRYs, while FBXL21, the closest paralog of FBXL3, ubiquitinates CRYs but leads to stabilization of CRYs. Fbxl3 knockout extremely lengthened the circadian period, and deletion of Fbxl21 gene in Fbxl3-deficient mice partially rescued the period-lengthening phenotype, suggesting a key role of CRY protein stability for maintenance of the circadian periodicity. Here, we employed a proteomics strategy to explore regulators for the protein stability of CRYs. We found that ubiquitin-specific protease 7 (USP7 also known as HAUSP) associates with CRY1 and CRY2 and stabilizes CRYs through deubiquitination. Treatment with USP7-specific inhibitor or Usp7 knockdown shortened the circadian period of the cellular rhythm. We identified another CRYs-interacting protein, TAR DNA binding protein 43 (TDP-43), an RNA-binding protein. TDP-43 stabilized CRY1 and CRY2, and its knockdown also shortened the circadian period in cultured cells. The present study identified USP7 and TDP-43 as the regulators of CRY1 and CRY2, underscoring the significance of the stability control process of CRY proteins for period determination in the mammalian circadian clockwork.