a Department of Biosciences & Bioengineering , Indian Institute of Technology Bombay , Mumbai , India.
Cell Cycle. 2018;17(16):2052-2068. doi: 10.1080/15384101.2018.1513764. Epub 2018 Sep 11.
Centrosomal proteins play important roles in the spindle assembly and the segregation of chromosomes in the eukaryotic cells. STARD9, a recently identified centrosomal protein, was reported to influence the spindle pole assembly. However, the role of STARD9 in maintaining the stability and organization of microtubules are not known. Here, we show that STARD9 regulates the assembly and dynamics of both interphase and mitotic microtubules. The knockdown of STARD9 in HeLa or HCT116 cells with siRNA or shRNA induced a strong depolymerization of the interphase microtubules. The over-expression of the motor domain of STARD9 stabilizes microtubules against cold and nocodazole suggesting that STARD9 stabilizes microtubules in HeLa cells. Using fluorescent recovery after photobleaching, we showed that the knockdown of STARD9 strongly reduced microtubule dynamics in the live spindles of HeLa cells. The reassembly of microtubules in the STARD9-depleted cells was strongly reduced as compared to the microtubules in the control cells implying the role of STARD9 in the nucleation of microtubules. Further, the depletion of STARD9 inhibited chromosome separation and the STARD9-depleted HeLa cells were blocked at mitosis. Interestingly, the frequency of multipolar spindle formation increased significantly in the STARD9-depleted HeLa cells in the presence of vinblastine and the STARD9-depleted cells showed much higher sensitivity towards vinblastine than the control cells indicating a new approach for cancer chemotherapy. The evidence suggests that STARD9 regulates the assembly and stability of both interphase and spindle microtubules and thereby, play important roles in the cell cycle progression.
中心体蛋白在真核细胞的纺锤体组装和染色体分离中发挥重要作用。STARD9 是一种新鉴定的中心体蛋白,据报道它会影响纺锤体极的组装。然而,STARD9 维持微管稳定性和组织的作用尚不清楚。在这里,我们发现 STARD9 调节间期和有丝分裂微管的组装和动态。用 siRNA 或 shRNA 敲低 HeLa 或 HCT116 细胞中的 STARD9 会诱导间期微管的强烈解聚。STARD9 马达结构域的过表达稳定了微管对冷和诺考达唑的作用,表明 STARD9 稳定了 HeLa 细胞中的微管。通过荧光恢复后漂白,我们表明 STARD9 的敲低强烈降低了活 HeLa 细胞纺锤体中微管的动力学。与对照细胞中的微管相比,STARD9 耗尽细胞中微管的重新组装大大减少,这意味着 STARD9 在微管的成核中起作用。此外,STARD9 的耗竭抑制了染色体分离,STARD9 耗尽的 HeLa 细胞在有丝分裂中被阻断。有趣的是,在长春花碱存在的情况下,STARD9 耗尽的 HeLa 细胞中多极纺锤体的形成频率显著增加,并且 STARD9 耗尽的细胞对长春花碱的敏感性比对照细胞高得多,这表明了癌症化疗的新方法。这些证据表明,STARD9 调节间期和纺锤体微管的组装和稳定性,从而在细胞周期进程中发挥重要作用。