Faculty of Medicine and Health Sciences, International University of Catalonia, 08195, Sant Cugat del Vallès, Barcelona, Spain.
Department of Cellular and Molecular Pharmacology and Howard Hughes Medical Institute, University of California, San Francisco, CA, 94158, USA.
Exp Mol Med. 2019 Apr 16;51(4):1-17. doi: 10.1038/s12276-019-0242-2.
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.
CDK16（也称为 PCTAIRE1 或 PCTK1）是细胞周期蛋白依赖性激酶（CDK）家族的一个非典型成员，它与细胞周期蛋白 Y（CCNY）形成一个活性复合物。尽管这两种蛋白质最近都被牵连到癌症的发病机制中，但 CDK16/CCNY 复合物如何发挥其生物学活性仍不清楚。为了了解 CDK16/CCNY 网络，我们使用互补的蛋白质组学方法来鉴定该复合物的潜在底物。我们鉴定了几个候选物，表明 CDK16/CCNY 复合物参与细胞骨架动力学，我们专注于微管相关蛋白有丝分裂调控因子（PRC1），它是细胞分裂所必需的蛋白质，组织对向微管，其失调可能导致癌症中的基因组不稳定。我们使用 CRISPR-Cas9 诱变在 293T 细胞中生成的模拟敏感（AS）CDK16，发现 CDK16 的特异性抑制诱导 PRC1 在 Threonine481 上的去磷酸化和在有丝分裂间期向核内的重新定位。CDK16 抑制和 PRC1 下调对细胞活力表现出上位性效应的观察结果证实，这些蛋白质可以通过单一途径发挥作用。总之，我们确定了 PRC1 是 CDK16/CCNY 复合物的第一个底物，并证明了 CDK16 的增殖功能是通过 PRC1 的磷酸化介导的。由于 CDK16 作为癌症的关键节点正在出现，我们的研究揭示了新的潜在治疗靶点。
