Spatiotemporal manipulation of ciliary glutamylation reveals its roles in intraciliary trafficking and Hedgehog signaling

• Published in: Nature communications Vol. 9; no. 1; pp. 1732 - 13
• Main Authors: Hong, Shi-Rong, Wang, Cuei-Ling, Huang, Yao-Shen, Chang, Yu-Chen, Chang, Ya-Chu, Pusapati, Ganesh V., Lin, Chun-Yu, Hsu, Ning, Cheng, Hsiao-Chi, Chiang, Yueh-Chen, Huang, Wei-En, Shaner, Nathan C., Rohatgi, Rajat, Inoue, Takanari, Lin, Yu-Chun
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30.04.2018; Nature Publishing Group; Nature Portfolio
• Subjects: 13/109; 14/1; 14/35; 14/63; 631/80/128/1383; 631/80/128/1653; Acetylation; Animals; Cell surface; Cilia; Cilia - metabolism; Cilia - ultrastructure; Crosstalk; Enzymes; Feasibility studies; Genes, Reporter; Genetic engineering; Glutamic Acid - metabolism; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Hedgehog protein; Hedgehog Proteins - genetics; Hedgehog Proteins - metabolism; Humanities and Social Sciences; Kinesin; Localization; Medicine; Mice; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; multidisciplinary; NIH 3T3 Cells; Optical Imaging; Post-translation; Protein Processing, Post-Translational; Proteins; Science; Science (multidisciplinary); Signal Transduction; Signaling; Tubulin; Tubulin - genetics; Tubulin - metabolism; Tyrosine - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-018-03952-z

Tubulin post-translational modifications (PTMs) occur spatiotemporally throughout cells and are suggested to be involved in a wide range of cellular activities. However, the complexity and dynamic distribution of tubulin PTMs within cells have hindered the understanding of their physiological roles in specific subcellular compartments. Here, we develop a method to rapidly deplete tubulin glutamylation inside the primary cilia, a microtubule-based sensory organelle protruding on the cell surface, by targeting an engineered deglutamylase to the cilia in minutes. This rapid deglutamylation quickly leads to altered ciliary functions such as kinesin-2-mediated anterograde intraflagellar transport and Hedgehog signaling, along with no apparent crosstalk to other PTMs such as acetylation and detyrosination. Our study offers a feasible approach to spatiotemporally manipulate tubulin PTMs in living cells. Future expansion of the repertoire of actuators that regulate PTMs may facilitate a comprehensive understanding of how diverse tubulin PTMs encode ciliary as well as cellular functions. Tubulin post-translational modifications (PTMs) occur spatiotemporally throughout cells, therefore assessing the physiological roles in specific subcellular compartments has been challenging. Here the authors develop a method to rapidly deplete tubulin glutamylation inside the primary cilia by targeting an engineered deglutamylase to the axoneme.