Global site-specific neddylation profiling reveals that NEDDylated cofilin regulates actin dynamics

Vogl, Annette M.; Phu, Lilian; Becerra, Raquel; Giusti, Sebastian A.; Verschueren, Erik; Hinkle, Trent B.; Bordenave, Martin D.; Adrian, Max; Heidersbach, Amy; Yankilevich, Patricio; Stefani, Fernando D.; Wurst, Wolfgang; Hoogenraad, Casper C.; Kirkpatrick, Donald S.; Refojo, Damian und Sheng, Morgan (2020): Global site-specific neddylation profiling reveals that NEDDylated cofilin regulates actin dynamics. In: Nature Structural & Molecular Biology, Bd. 27, Nr. 2

Abstract

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-epsilon GG-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.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin > Munich Cluster for Systems Neurology (SyNergy)
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• Sprache: Englisch
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 390857198