A.N. Belozersky Institute for Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Vorobjevy Gory, 1 bld.73, Moscow, Russian Federation, 119991.
Department of Cell Biology of Institute of Protein Research, Russian Academy of Science, Vavilova ul., 34, Moscow, Russian Federation, 117334.
Protoplasma. 2019 Sep;256(5):1361-1373. doi: 10.1007/s00709-019-01385-z. Epub 2019 May 11.
In animal cells, the centrosome nucleates and anchors microtubules (MT), forming their radial array. During interphase centrosome-derived MT, aster can either team up with other MT network or function in an autonomous manner. What is the function of the centrosome-derived MT aster? We suggested that it might play an important role in the formation of the primary cilium, the organelle obligatorily associated with the centrosome. PCM-1 (PeriCentriolar Matrix 1) protein, which participates in the organization of the primary cilium, is a part of pericentiolar satellites. They are transported to the centrosome along MTs by the motor protein dynein in a complex with its cofactor dynactin. Previously, we showed that SLK/LOSK phosphorylated the p150 subunit of dynactin, thus promoting its centrosomal targeting followed by its participation in the retention of microtubules. Here, we found that under the repression of SLK/LOSK activity, the PCM-1 protein lost its pericentrosomal localization and was being dispersed throughout the cytoplasm. Despite that the alanine and glutamine mutants of p150 had opposite effects on PCM-1 localization, they associated with PCM-1 to the same extent. The occurrence of primary cilia also significantly decreased when SLK/LOSK was repressed. These defects also correlated with a disturbance of the long-range transport in cells, whereas dynein-depending motility was intact. Treatment with the GSK-3β kinase inhibitor also resulted in the loss of the centrosome-derived MT aster, dispersion of PCM-1 over the cytoplasm, and reduction of primary cilia occurrence. Thus, kinases involved in the centrosome-derived MT aster regulation can indirectly control the formation of primary cilia in cells.
在动物细胞中,中心体起始并锚定微管(MT),形成其放射状排列。在间期,中心体衍生的 MT 星体可以与其他 MT 网络结合,或者以自主的方式发挥作用。那么,中心体衍生的 MT 星体的功能是什么呢?我们推测,它可能在初级纤毛的形成中发挥重要作用,而初级纤毛是与中心体必然相关的细胞器。参与初级纤毛组织的 PCM-1(PeriCentriolar Matrix 1)蛋白是中心粒卫星的一部分。它们通过与动力蛋白 dynein 及其共因子 dynactin 形成的复合物沿 MT 被运输到中心体。此前,我们表明 SLK/LOSK 磷酸化 dynactin 的 p150 亚基,从而促进其向中心体的靶向,随后参与微管的保留。在这里,我们发现在 SLK/LOSK 活性受到抑制的情况下,PCM-1 蛋白失去了中心体周围的定位,并分散在细胞质中。尽管 p150 的丙氨酸和谷氨酰胺突变体对 PCM-1 定位有相反的影响,但它们与 PCM-1 的结合程度相同。当 SLK/LOSK 受到抑制时,初级纤毛的发生也显著减少。这些缺陷也与细胞内长距离运输的干扰相关,而依赖 dynein 的运动是完整的。用 GSK-3β 激酶抑制剂处理也会导致中心体衍生的 MT 星体丢失、PCM-1 在细胞质中弥散以及初级纤毛发生减少。因此,参与中心体衍生的 MT 星体调节的激酶可以间接控制细胞中初级纤毛的形成。