Comparative Proteomics Reveals Timely Transport into Cilia of Regulators or Effectors as a Mechanism Underlying Ciliary Disassembly
Comparative Proteomics Reveals Timely Transport into Cilia of Regulators or Effectors as a Mechanism Underlying Ciliary Disassembly
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比较蛋白质组学揭示了调节器或效应器及时转运到纤毛中作为纤毛分解的机制
DOI：
10.1021/acs.jproteome.6b01048
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发表时间：
2017
影响因子：
4.4
通讯作者：
Pan Junmin
中科院分区：
文献类型：
--
作者：
Wang Limei;Gu Lixiao;Meng Dan;Wu Qiong;Deng Haiteng;Pan Junmin
初级纤毛在细胞周期过程中组装和拆卸。在纤毛拆卸过程中，纤毛轴突微管（MTs）被解聚，同时伴随着纤毛蛋白的广泛翻译后蛋白修饰，包括蛋白磷酸化、甲基化和泛素化。假设这些事件涉及在纤毛从细胞体拆卸时将效应器或调节剂运输到纤毛中。为了证明这一假设，并发现参与纤毛拆卸的新蛋白质，我们使用比较蛋白质组学和TMT标记分析了衣原体鞭毛的拆卸。在四个重复中发现91个蛋白质增加了1.4倍以上。IFT机制的蛋白质不仅增加，而且表现出化学计量的变化。其他增加的蛋白质包括信号分子、伴侣蛋白和参与微管动力学或稳定性的蛋白质。特别是，我们已经确定了一种纤毛病蛋白C21orf2，即aaa - atp酶CDC48，它参与从大组件或细胞结构中分离多肽，FAP203和FAP236，它们是轴突微管稳定剂的同源物。我们的数据表明，纤毛转运效应器或调节器是潜在的纤毛拆卸机制之一。进一步表征所鉴定的蛋白质将为我们对纤毛拆卸和可能的纤毛病的理解提供新的见解。
Primary cilia are assembled and disassembled during cell cycle progression. During ciliary disassembly, ciliary axonemal microtubules (MTs) are depolymerized accompanied by extensive posttranslational protein modifications of ciliary proteins including protein phosphorylation, methylation, and ubiquitination. These events are hypothesized to involve transport of effectors or regulators into cilia at the time of ciliary disassembly from the cell body. To prove this hypothesis and identify new proteins involved in ciliary disassembly, we analyzed disassembling flagella inChlamydomonasusing comparative proteomics with TMT labeling. Ninety-one proteins were found to increase more than 1.4-fold in four replicates. The proteins of the IFT machinery not only increase but also exhibit stoichiometric changes. The other proteins that increase include signaling molecules, chaperones, and proteins involved in microtubule dynamics or stability. In particular, we have identified a ciliopathy protein C21orf2, the AAA-ATPase CDC48, that is involved in segregating polypeptides from large assemblies or cellular structures, FAP203 and FAP236, which are homologous to stabilizers of axonemal microtubules. Our data demonstrate that ciliary transport of effectors or regulators is one of the mechanisms underlying ciliary disassembly. Further characterization of the proteins identified will provide new insights into our understanding of ciliary disassembly and likely ciliopathy.