Global site-specific neddylation profiling reveals that NEDDylated cofilin regulates actin dynamics
| Autor: | Vogl, Annette M., Phu, Lilian, Becerra, Raquel, Giusti, Sebastian A., Verschueren, Erik, Hinkle, Trent B., Bordenave, Martín D., Adrian, Max, Heidersbach, Amy, Yankilevich, Patricio, Stefani, Fernando D., Wurst, Wolfgang, Hoogenraad, Casper C., Kirkpatrick, Donald S., Refojo, Damian, Sheng, Morgan, Celbiologie, Sub Cell Biology |
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| Přispěvatelé: | Celbiologie, Sub Cell Biology |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Cofilin 1
Neurite metabolism [Actins] NEDD8 Protein Lysine metabolism [NEDD8 Protein] macromolecular substances NEDD8 Cell Line Rats Sprague-Dawley 03 medical and health sciences Mice 0302 clinical medicine Ubiquitin Structural Biology ddc:570 metabolism [Ubiquitin] Animals Humans Point Mutation Gene Knock-In Techniques Cytoskeleton Molecular Biology Actin Cells Cultured 030304 developmental biology Neurons 0303 health sciences biology Chemistry Ubiquitination Cofilin Actins Cell biology Rats metabolism [Cofilin 1] Mice Inbred C57BL HEK293 Cells metabolism [Neurons] genetics [NEDD8 Protein] cytology [Neurons] biology.protein Neddylation 030217 neurology & neurosurgery |
| Zdroj: | Nature structural & molecular biology 27(2), 210-220 (2020). doi:10.1038/s41594-019-0370-3 Nature Structural and Molecular Biology, 27(2), 210. Nature Publishing Group |
| ISSN: | 1545-9993 |
| DOI: | 10.1038/s41594-019-0370-3 |
| Popis: | Neddylation is the post-translational protein modification most closely related to ubiquitination. Whereas the ubiquitin-like protein NEDD8 is well studied for its role in activating cullin−RING E3 ubiquitin ligases, little is known about other substrates. We developed serial NEDD8-ubiquitin substrate profiling (sNUSP), a method that employs NEDD8 R74K knock-in HEK293 cells, allowing discrimination of endogenous NEDD8- and ubiquitin-modification sites by MS after Lys-C digestion and K-eGG-peptide enrichment. Using sNUSP, we identified 607 neddylation sites dynamically regulated by the neddylation inhibitor MLN4924 and the de-neddylating enzyme NEDP1, implying that many non-cullin proteins are neddylated. Among the candidates, we characterized lysine 112 of the actin regulator cofilin as a novel neddylation event. Global inhibition of neddylation in developing neurons leads to cytoskeletal defects, altered actin dynamics and neurite growth impairments, whereas site-specific neddylation of cofilin at K112 regulates neurite outgrowth, suggesting that cofilin neddylation contributes to the regulation of neuronal actin organization. NEDD8-ubiquitin substrate profiling (sNUSP) identifies neddylation sites in many non-cullin proteins. Among the candidates, neddylation of cofilin regulates actin dynamics and neurite growth and outgrowth in developing neurons. |
| Databáze: | OpenAIRE |
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