Various effects of two types of kinesin-5 inhibitors on mitosis and cell proliferation
Various effects of two types of kinesin-5 inhibitors on mitosis and cell proliferation
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DOI：
10.1016/j.bcp.2021.114789
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发表时间：
2021-10-07
影响因子：
5.8
通讯作者：
Asai，Akira
中科院分区：
文献类型：
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作者：
Sawada，Jun-ichi;Matsuno，Kenji;Asai，Akira
驱动蛋白-5作为有丝分裂的新靶点受到了广泛关注。在过去的几十年中，已经开发了各种靶向驱动蛋白-5的小分子化合物。然而，驱动蛋白-5抑制剂的细胞效应的差异仍然知之甚少。在这里，我们使用了两种不同的驱动蛋白-5抑制剂，联苯型PVZB 1194和S-三苯甲基-L-半胱氨酸型PVEI 0021，以检查它们对涉及驱动蛋白-5的分子事件的影响。我们对驱动蛋白-5蛋白-蛋白相互作用的生化研究表明，PVZB 1194处理的驱动蛋白-5与TPX 2微管成核因子、Aurora-A激酶、透明质酸介导的运动受体和γ-微管蛋白相互作用，与未处理的有丝分裂驱动蛋白-5一样。然而，PVEI 0021阻止驱动蛋白-5与这些蛋白质结合。在从诺考达唑抑制中恢复的有丝分裂HeLa细胞中，驱动蛋白-5与这些结合蛋白共定位，沿着与着丝粒附近的微管成核。PVZB 1194通过影响驱动蛋白5与染色质相关微管的相互作用，而不是PVEI 0021，不仅影响了分散的微管簇的形成，而且增强了微管的稳定性。此外，筛选与驱动蛋白-5抑制剂协同作用的有丝分裂抑制剂表明，紫杉醇仅与PVZB 1194协同抑制HeLa细胞增殖。相比之下，Aurora-A抑制剂MLN 8237与任一抑制剂组合时发挥协同抗细胞增殖作用。总之，这些结果提供了一个更好的理解驱动蛋白-5抑制剂的分子作用，并表明其有用的有丝分裂的研究和抗癌药物的发展作为分子工具。
Kinesin-5 has received considerable attention as a new target for mitosis. Various small-molecule compounds targeting kinesin-5 have been developed in the last few decades. However, the differences in the cellular effects of kinesin-5 inhibitors remain poorly understood. Here, we used two different kinesin-5 inhibitors, biphenyl-type PVZB1194 andS‐trityl‐L‐cysteine-type PVEI0021, to examine their effects on molecular events involving kinesin-5. Our biochemical study of kinesin-5 protein-protein interactions showed that PVZB1194-treated kinesin-5 interacted with TPX2 microtubule nucleation factor, Aurora-A kinase, receptor for hyaluronan-mediated motility, and γ-tubulin, as did untreated mitotic kinesin-5. However, PVEI0021 prevented kinesin-5 from binding to these proteins. In mitotic HeLa cells recovered from nocodazole inhibition, kinesin-5 colocalized with these binding proteins, along with microtubules nucleated near kinetochores. By acting on kinesin-5 interactions with chromatin-associated microtubules, PVZB1194, rather than PVEI0021, not only affected the formation of dispersed microtubule clusters but also enhanced the stability of microtubules. In addition, screening for mitotic inhibitors working synergistically with the kinesin-5 inhibitors revealed that paclitaxel synergistically inhibited HeLa cell proliferation only with PVZB1194. In contrast, the Aurora-A inhibitor MLN8237 exerted a synergistic anti-cell proliferation effect when combined with either inhibitor. Together, these results have provided a better understanding of the molecular action of kinesin-5 inhibitors and indicate their usefulness as molecular tools for the study of mitosis and the development of anticancer agents.