10.1021/acs.jproteome.5b00308.s002 Ningning Liu Ningning Liu Yun Xiong Yun Xiong Yiran Ren Yiran Ren Linlin Zhang Linlin Zhang Xianfei He Xianfei He Xincheng Wang Xincheng Wang Min Liu Min Liu Dengwen Li Dengwen Li Wenqing Shui Wenqing Shui Jun Zhou Jun Zhou Proteomic Profiling and Functional Characterization of Multiple Post-Translational Modifications of Tubulin American Chemical Society 2015 tubulin PTMs CTT microtubule role tubulin phosphorylation 24 novel PTM sites 2015-08-07 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/Proteomic_Profiling_and_Functional_Characterization_of_Multiple_Post_Translational_Modifications_of_Tubulin/2143306 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 <i>O</i>-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.