Phosphoregulation of the oncogenic protein regulator of cytokinesis 1 (PRC1) by the atypical CDK16/CCNY complex.
Phosphoregulation of the oncogenic protein regulator of cytokinesis 1 (PRC1) by the atypical CDK16/CCNY complex.
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DOI：
10.1038/s12276-019-0242-2
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发表时间：
2019-04-16
影响因子：
12.8
通讯作者：
Clotet, Josep
中科院分区：
文献类型：
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作者：
Hernandez-Ortega, Sara;Sanchez-Botet, Abril;Clotet, Josep
CDK16(又称PCTAIRE1或PCTK1)是细胞周期蛋白依赖性蛋白激酶(CDK)家族中的一个非典型成员，与细胞周期蛋白Y(CCNY)形成活性复合体。虽然这两种蛋白最近都与癌症的发病机制有关，但CDK16/CCNY复合体如何发挥其生物活性仍不清楚。为了了解CDK16/CCNY网络，我们使用互补蛋白质组学方法来确定该复合体的潜在底物。我们确定了几个与细胞骨架动力学有关的CDK16/CCNY复合体的候选基因，并重点研究了胞质微管相关蛋白调节因子(PRC1)，这是一种对细胞分裂至关重要的蛋白质，它组织反平行的微管，其调控的解除可能导致癌症基因组的不稳定。在293T细胞中使用CRISPR-Cas9突变产生的模拟敏感(AS)CDK16，我们发现CDK16的特异性抑制诱导了Prc1在Thr481处的去磷酸化并在间期引起核离位。观察到CDK16抑制和PRC1下调对细胞活力具有上位性效应，证实了这些蛋白可以通过单一途径发挥作用。综上所述，我们确定Prc1是CDK16/CCNY复合体的第一底物，并证明CDK16的增殖功能是通过Prc1磷酸化来实现的。由于CDK16正在成为癌症的关键节点，我们的研究揭示了新的潜在治疗靶点。
CDK16 (also known as PCTAIRE1 or PCTK1) is an atypical member of the cyclin-dependent kinase (CDK) family that forms an active complex with cyclin Y (CCNY). Although both proteins have been recently implicated in cancer pathogenesis, it is still unclear how the CDK16/CCNY complex exerts its biological activity. To understand the CDK16/CCNY network, we used complementary proteomic approaches to identify potential substrates of this complex. We identified several candidates implicating the CDK16/CCNY complex in cytoskeletal dynamics, and we focused on the microtubule-associated protein regulator of cytokinesis (PRC1), an essential protein for cell division that organizes antiparallel microtubules and whose deregulation may drive genomic instability in cancer. Using analog-sensitive (AS) CDK16 generated by CRISPR-Cas9 mutagenesis in 293T cells, we found that specific inhibition of CDK16 induces PRC1 dephosphorylation at Thr481 and delocalization to the nucleus during interphase. The observation that CDK16 inhibition and PRC1 downregulation exhibit epistatic effects on cell viability confirms that these proteins can act through a single pathway. In conclusion, we identified PRC1 as the first substrate of the CDK16/CCNY complex and demonstrated that the proliferative function of CDK16 is mediated by PRC1 phosphorylation. As CDK16 is emerging as a critical node in cancer, our study reveals novel potential therapeutic targets.