Proteomic Profiling and Functional Characterization of Multiple Post-Translational Modifications of Tubulin
Proteomic Profiling and Functional Characterization of Multiple Post-Translational Modifications of Tubulin
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微管蛋白多种翻译后修饰的蛋白质组分析和功能表征
DOI：
10.1021/acs.jproteome.5b00308
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发表时间：
2015-08-01
影响因子：
4.4
通讯作者：
Zhou, Jun
中科院分区：
文献类型：
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作者：
Liu, Ningning;Xiong, Yun;Zhou, Jun
众所周知，微管蛋白经历了独特的翻译后修饰(PTM)，如去酪氨酸化和多谷氨酰化，特别是在非结构化羧基末端尾巴(CTTS)。然而，更传统的微管蛋白PTMS及其在微管特性和功能调节中的作用仍然不清楚。在这里，我们报道了用蛋白质组学的方法对HeLa细胞中微管蛋白的磷酸化、乙酰化、泛素化和O-GlcN酰化进行了全面的研究。我们的微管蛋白靶向分析已经确定了80个残基携带单个或多个传统的PTM，其中包括24个新的PTM位点，这在以前的全球蛋白质组研究中没有涵盖。通过使用一系列PTM缺失或模仿PTM的突变体，我们进一步发现微管蛋白的磷酸化和乙酰化在控制微管组装和稳定性方面发挥着重要作用。此外，这些微管蛋白PTM对腺病毒沿微管的逆行运输具有明显的作用。因此，这些发现扩大了微管蛋白PTM的谱系，并促进了我们对它们在微管动力学和细胞功能调节中的多种作用的理解。
Tubulin is known to undergo unique post-translational modifications (PTMs), such as detyrosination and polyglutamylation, particularly in the unstructured carboxy-terminal tails (CTTs). However, more conventional PTMs of tubulin and their roles in the regulation of microtubule properties and functions remain poorly defined. Here, we report the comprehensive profiling of tubulin phosphorylation, acetylation, ubiquitylation, and O-GlcNAcylation in HeLa cells with a proteomic approach. Our tubulin-targeted analysis has identified 80 residues bearing single or multiple conventional PTMs including 24 novel PTM sites not covered in previous global proteomic surveys. By using a series of PTM-deficient or PTM-mimicking mutants, we further find that tubulin phosphorylation and acetylation play important roles in the control of microtubule assembly and stability. In addition, these tubulin PTMs have distinct effects on the retrograde transport of adenoviruses along microtubules. These findings thus enlarge the repertoire of tubulin PTMs and foster our understanding of their versatile roles in the regulation of microtubule dynamics and cellular functions.