Subdiffraction-resolution fluorescence microscopy reveals a domain of the centrosome critical for pericentriolar material organization.

Subdiffraction-resolution fluorescence microscopy reveals a domain of the centrosome critical for pericentriolar material organization.
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DOI:
10.1038/ncb2597
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发表时间:
2012-11
影响因子:
21.3
通讯作者:
--
中科院分区:
生物学1区
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--
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中心体作为动物细胞中微管的主要组织中心,在细胞功能中起着基础性作用。围绕中心粒的周中心粒物质(Pericentriolar material,PCM)为微管成核提供了动态平台。虽然PCM的重要性已经确立,但其非晶电子致密的性质使其难以进行结构研究。通过使用SIM和STORM亚衍射分辨率显微镜来可视化中心体成熟的关键蛋白质,我们证明了PCM被组织成两个主要的结构域:与中心粒壁并列的一层,以及远离中心粒组织在基质中的蛋白质。对Pericentrin样蛋白(Plp)的分析表明,它的C-末端位于中心粒壁,它向外辐射到基质中,并组织成具有准九重对称性的簇。通过RNAi,我们表明,Plp纤维所需的间期招聘和适当的有丝分裂组装的PCM矩阵。
As the main microtubule-organizing center in animal cells, the centrosome has a fundamental role in cell function. Surrounding the centrioles, the Pericentriolar material (PCM) provides a dynamic platform for nucleating microtubules. While the PCM’s importance is established, its amorphous electron-dense nature has made it refractory to structural investigation. By using SIM and STORM sub-diffraction resolution microscopies to visualize proteins critical for centrosome maturation, we demonstrate that the PCM is organized into two major structural domains: a layer juxtaposed to the centriole wall, and proteins extending further away from the centriole organized in a matrix. Analysis of Pericentrin-like protein (Plp) reveals that its C-terminus is positioned at the centriole wall, it radiates outward into the matrix and is organized in clusters having quasi-nine-fold symmetry. By RNAi we show that Plp fibrils are required for interphase recruitment and proper mitotic assembly of the PCM matrix.
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