CDK5RAP3, a Novel Nucleoplasmic Shuttle, Deeply Regulates HSF1-Mediated Heat Stress Response and Protects Mammary Epithelial Cells from Heat Injury.
CDK5RAP3, a Novel Nucleoplasmic Shuttle, Deeply Regulates HSF1-Mediated Heat Stress Response and Protects Mammary Epithelial Cells from Heat Injury.
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CDK5RAP3 是一种新型核质梭，深度调节 HSF1 介导的热应激反应并保护乳腺上皮细胞免受热损伤
DOI：
10.3390/ijms21218400
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发表时间：
2020-11-09
影响因子：
5.6
通讯作者：
Cai Y
中科院分区：
文献类型：
--
作者：
Shen Y;Zou Y;Li J;Chen F;Li H;Cai Y
CDK 5 RAP 3被认为是对抗热应激的细胞应答的最重要的调节剂，热应激与免疫系统的功能障碍和动物对疾病的易感性有关。尽管如此，关于CDK 5 RAP 3如何调节热应激反应的研究却知之甚少。本研究分别以CDK 5 RAP 3条件性基因敲除（CKO）小鼠、CDK 5 RAP 3-/-小鼠胚胎成纤维细胞（MEFs）和牛乳腺上皮细胞（BMEC）为体内外模型，揭示CDK 5 RAP 3在热应激反应中的调控作用。CDK 5 RAP 3的缺失在1.5小时热刺激后意外地导致动物死亡。热应激8h后再培养BMEC，CDK 5 RAP 3和HSPs的表达均呈上升（0-2、4-6 h）和下降（2-4、6- 8h）的波动规律。CDK 5 RAP 3的缺失除了显著增强热休克蛋白的表达、凋亡率和内质网应激外，还影响核质转位和热休克因子1（HSF 1）三聚体的形成。这些程序也在CDK 5 RAP 3 CKO小鼠和CDK 5 RAP 3-/- MEFs的乳腺中得到进一步证实。有趣的是，HSF 1基因沉默下调了BMEC中CDK 5 RAP 3的表达。免疫染色和免疫沉淀研究表明CDK 5 RAP 3和HSF 1之间的物理相互作用共定位于细胞质和细胞核中。此外，CDK 5 RAP 3还与HSP 90相互作用，这表明在转录水平和HSP 90本身的蛋白质功能上都有一个操作机制。总之，我们的研究结果表明，CDK 5 RAP 3像一种新的核质穿梭或分子伴侣一样工作，深入参与HSF 1介导的热应激反应，保护细胞免受热损伤。
CDK5RAP3 was regarded as the most significant regulator of cellular responses against heat stress, which is associated with dysfunctions of the immune system and animal susceptibility to disease. Despite this, little known about how CDK5RAP3 regulates heat stress response. In this study, CDK5RAP3 conditional Knockout (CKO) mice, CDK5RAP3-/- mouse embryo fibroblasts (MEFs) and bovine mammary epithelial cells (BMECs) were used as an in vitro and in vivo model, respectively to reveal the role of CDK5RAP3 in regulating the heat stress response. The deletion of CDK5RAP3 unexpectedly caused animal lethality after 1.5-h heat stimulations. Furthermore, BMECs were re-cultured for eight hours after heat stress and was found that the expression of CDK5RAP3 and HSPs showed a similar fluctuating pattern of increase (0–2, 4–6 h) and decrease (2–4, 6–8 h). In addition to the remarkably enhanced expression of heat shock protein, apoptosis rate and endoplasmic reticulum stress, the deletion of CDK5RAP3 also affected nucleoplasmic translocation and trimer formation of heat shock factor 1 (HSF1). These programs were further confirmed in the mammary gland of CDK5RAP3 CKO mice and CDK5RAP3-/- MEFs as well. Interestingly, genetic silencing of HSF1 downregulated CDK5RAP3 expression in BMECs. Immunostaining and immunoprecipitation studies suggested a physical interaction between CDK5RAP3 and HSF1 being co-localized in the cytoplasm and nucleus. Besides, CDK5RAP3 also interacted with HSP90, suggesting an operative machinery at both transcriptional level and protein functionality of HSP90 per se. Together, our findings suggested that CDK5RAP3 works like a novel nucleoplasmic shuttle or molecular chaperone, deeply participating in HSF1-mediated heat stress response and protecting cells from heat injury.
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DOI：
10.1007/978-3-030-40204-4_3
发表时间：
2020-01-01
期刊：
HSF1 AND MOLECULAR CHAPERONES IN BIOLOGY AND CANCER
影响因子：
--
作者：
Pincus, David
通讯作者：
Pincus, David
影响因子：
5.3
作者：
Budzynski, Marek A.;Puustinen, Mikael C.;Sistonen, Lea
通讯作者：
Sistonen, Lea
影响因子：
2.7
作者：
Bhanuprakash, V.;Singh, Umesh;Deb, Rajib
通讯作者：
Deb, Rajib
影响因子：
3.8
作者：
Cai, Mingcheng;Hu, Yongsong;Lai, Songjia
通讯作者：
Lai, Songjia
影响因子：
11.2
作者：
Mak, Grace Wing-Yan;Chan, Mandy Man-Lok;Ching, Yick-Pang
通讯作者：
Ching, Yick-Pang