Protein quality control of DYRK family protein kinases by the Hsp90-Cdc37 molecular chaperone.
Protein quality control of DYRK family protein kinases by the Hsp90-Cdc37 molecular chaperone.
复制标题
Hsp90-Cdc37 分子伴侣对 DYRK 家族蛋白激酶的蛋白质质量控制。
DOI：
10.1016/j.bbamcr.2021.119081
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
E.
中科院分区：
文献类型：
--
作者：
Miyata;Y.;and Nishida;E.
DYRK（双特异性酪氨酸磷酸化调节蛋白激酶）家族由五种相关蛋白激酶（DYRK1A、DYRK1B、DYRK2、DYRK3、DYRK4）组成。 DYRK 与果蝇小脑具有同源性，人类 21 号染色体上的 DYRK1A 与包括人类唐氏综合症在内的各种神经元疾病有关。在这里，我们报告了与 DYRK 特定成员相关的细胞蛋白的鉴定。与 DYRK1B 和 DYRK4 相关的分子量为 90、70 和 50 kDa 的细胞蛋白。这些蛋白分别被鉴定为分子伴侣 Hsp90、Hsp70 和 Cdc37。 GFP-DYRK 的显微镜分析显示，在 COS7 细胞中，DYRK1A 和 DYRK1B 位于细胞核，而 DYRK2、DYRK3 和 DYRK4 大部分位于细胞质。 DYRK1B 的过度表达诱导这些分子伴侣与 DYRK1B 一起重新定位于核。用特异性 Hsp90 抑制剂格尔德霉素和 17-AAG 处理细胞，消除了 Hsp90 和 Cdc37 与 DYRK1B 和 DYRK4 的关联，但不能消除 Hsp70 的关联。 Hsp90 伴侣活性的抑制影响 DYRK1B 和 DYRK4 的细胞内动态。格尔德霉素处理后，DYRK1B 和 DYRK4 快速形成细胞质点状点，表明 Hsp90 的伴侣功能是防止靶激酶蛋白聚集所必需的。格尔德霉素、17-AAG 或 ganetespib 对 Hsp90 的长期抑制会降低 DYRK1B 和 DYRK4 的细胞水平。最后，DYRK1B和DYRK4在细胞中被泛素化，并且通过格尔德霉素抑制Hsp90进一步增加泛素化的DYRK1B和DYRK4。综上所述，这些结果表明 Hsp90 和 Cdc37 能够区分 DYRK 激酶家族的特定成员，并在细胞中这些客户激酶的质量控制中发挥重要作用。
The DYRK (Dual-specificity tYrosine-phosphorylation Regulated protein Kinase) family consists of five related protein kinases (DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4). DYRKs show homology toDrosophila Minibrain, and DYRK1A in human chromosome 21 is responsible for various neuronal disorders including human Down syndrome. Here we report identification of cellular proteins that associate with specific members of DYRKs. Cellular proteins with molecular masses of 90, 70, and 50-kDa associated with DYRK1B and DYRK4. These proteins were identified as molecular chaperones Hsp90, Hsp70, and Cdc37, respectively. Microscopic analysis of GFP-DYRKs showed that DYRK1A and DYRK1B were nuclear, while DYRK2, DYRK3, and DYRK4 were mostly cytoplasmic in COS7 cells. Overexpression of DYRK1B induced nuclear re-localization of these chaperones with DYRK1B. Treatment of cells with specific Hsp90 inhibitors, geldanamycin and 17-AAG, abolished the association of Hsp90 and Cdc37 with DYRK1B and DYRK4, but not of Hsp70. Inhibition of Hsp90 chaperone activity affected intracellular dynamics of DYRK1B and DYRK4. DYRK1B and DYRK4 underwent rapid formation of cytoplasmic punctate dots after the geldanamycin treatment, suggesting that the chaperone function of Hsp90 is required for prevention of protein aggregation of the target kinases. Prolonged inhibition of Hsp90 by geldanamycin, 17-AAG, or ganetespib, decreased cellular levels of DYRK1B and DYRK4. Finally, DYRK1B and DYRK4 were ubiquitinated in cells, and ubiquitinated DYRK1B and DYRK4 further increased by Hsp90 inhibition with geldanamycin. Taken together, these results indicate that Hsp90 and Cdc37 discriminate specific members of the DYRK kinase family and play an important role in quality control of these client kinases in cells.