Laboratório Nacional de Biociências, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brazil.
J Proteome Res. 2010 Dec 3;9(12):6298-316. doi: 10.1021/pr100562w. Epub 2010 Oct 22.
Physical protein-protein interactions are fundamental to all biological processes and are organized in complex networks. One branch of the kinome network is the evolutionarily conserved NIMA-related serine/threonine kinases (Neks). Most of the 11 mammalian Neks studied so far are related to cell cycle regulation, and due to association with diverse human pathologies, Neks are promising chemotherapeutic targets. Human Nek6 was associated to carcinogenesis, but its interacting partners and signaling pathways remain elusive. Here we introduce hNek6 as a highly connected member in the human kinase interactome. In a more global context, we performed a broad data bank comparison based on degree distribution analysis and found that the human kinome is enriched in hubs. Our networks include a broad set of novel hNek6 interactors as identified by our yeast two-hybrid screens classified into 18 functional categories. All of the tested interactions were confirmed, and the majority of tested substrates were phosphorylated in vitro by hNek6. Notably, we found that hNek6 N-terminal is important to mediate the interactions with its partners. Some novel interactors also colocalized with hNek6 and γ-tubulin in human cells, pointing to a possible centrosomal interaction. The interacting proteins link hNek6 to novel pathways, for example, Notch signaling and actin cytoskeleton regulation, or give new insights on how hNek6 may regulate previously proposed pathways such as cell cycle regulation, DNA repair response, and NF-κB signaling. Our findings open new perspectives in the study of hNek6 role in cancer by analyzing its novel interactions in specific pathways in tumor cells, which may provide important implications for drug design and cancer therapy.
物理蛋白质-蛋白质相互作用是所有生物过程的基础，并组织在复杂的网络中。激酶组网络的一个分支是进化保守的 NIMA 相关丝氨酸/苏氨酸激酶 (Neks)。迄今为止研究的 11 种哺乳动物 Nek 中，大多数与细胞周期调控有关，由于与多种人类病理有关，Neks 是有前途的化疗靶点。人类 Nek6 与致癌作用有关，但它的相互作用伙伴和信号通路仍然难以捉摸。在这里，我们将 hNek6 介绍为人类激酶相互作用组中的一个高度连接的成员。在更广泛的背景下，我们基于度分布分析进行了广泛的数据库比较，发现人类激酶组富含枢纽。我们的网络包括通过酵母双杂交筛选鉴定的广泛的新型 hNek6 相互作用物，这些相互作用物分为 18 个功能类别。所有测试的相互作用都得到了确认，并且大多数测试的底物在体外都被 hNek6 磷酸化。值得注意的是，我们发现 hNek6 的 N 端对于介导与其伙伴的相互作用很重要。一些新的相互作用物也与人 hNek6 和γ-微管蛋白在人类细胞中共定位，指向可能的中心体相互作用。相互作用蛋白将 hNek6 与新的途径联系起来，例如 Notch 信号转导和肌动蛋白细胞骨架调节，或为 hNek6 如何调节先前提出的途径（如细胞周期调控、DNA 修复反应和 NF-κB 信号转导）提供新的见解。我们的研究结果通过分析 hNek6 在肿瘤细胞中特定途径的新相互作用，为癌症中 hNek6 作用的研究开辟了新的视角，这可能为药物设计和癌症治疗提供重要的启示。
