Cep128 associates with Odf2 to form the subdistal appendage of the centriole
Cep128 associates with Odf2 to form the subdistal appendage of the centriole
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DOI：
10.1111/gtc.12668
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发表时间：
2019-03-01
期刊：
影响因子：
2.1
通讯作者：
Tsukita, Sachiko
中科院分区：
文献类型：
--
作者：
Kashihara, Hiroka;Chiba, Shuhei;Tsukita, Sachiko
细胞中的母亲中心粒有两个附属物，远侧附属物（DA）和近远侧附属物（SDA），它们分别具有产生纤毛和组织细胞微管网络的作用。在Odf 2的敲除（KO）细胞中，DA和SDA的组分，两个附属物同时消失。然而，DA和SDA在OM下游独立形成但彼此接近的分子机制尚不清楚。在这里，使用超分辨率结构照明显微镜（SR-SIM），我们发现GFP标记的Odf 2的信号与免疫荧光标记的Cep 128的信号相当重叠。我们进一步发现，Cep 128敲除（KD）导致其他SDA组分从中心粒解离，包括中心素，Ndell，ninein和Cep 170，而OM仍然与中心粒相关。与此相反，DA组分保持与Cep 128 KD细胞的中心粒。与这一观察结果一致，我们确定Cep 128作为ODF 2相互作用蛋白的免疫沉淀。考虑到Cep 128缺失降低了中心粒微管的稳定性，我们的研究结果表明，Cep 128与Od 12在SDA组件的分级组装中引起微管组织功能。
The mother centriole in a cell has two appendages, the distal appendage (DA) and subdistal appendage (SDA), which have roles in generating cilia and organizing the cellular microtubular network, respectively. In the knockout (KO) cells of Odf2, the component of the DA and SDA, both appendages simultaneously disappear. However, the molecular mechanisms by which the DA and SDA form independently but close to each other downstream of OM are unknown. Here, using super-resolution structured illumination microscopy (SR-SIM), we found that the signal for GFP-tagged Odf2 overlapped considerably with that of immunofluorescently labeled Cep128. We further found that Cep128 knockdown (KD) caused the dissociation of other SDA components from the centriole, including centriolin, Ndell, ninein and Cep170, whereas OM was still associated with the centriole. In contrast, the DA components remained associated with the centriole in Cep128 KD cells. Consistent with this observation, we identified Cep128 as an Odf2-interacting protein by immunoprecipitation. Taken with the finding that Cep128 deletion decreased the stability of centriolar microtuhules, our results indicate that Cep128 associates with Od12 in the hierarchical assembly of SDA components to elicit the microtubule-organizing function.