Global Site-specific Neddylation Profiling Reveals That NEDDylated Cofilin Regulates Actin Dynamics

Overview

Journal Nat Struct Mol Biol

Specialties Cell Biology
Molecular Biology

Date 2020 Feb 5

PMID 32015554

Citations 44

Authors

Annette M Vogl

Lilian Phu

Raquel Becerra

Sebastian A Giusti

Erik Verschueren

Trent B Hinkle

Martin D Bordenave

Max Adrian

Amy Heidersbach

Patricio Yankilevich

Fernando D Stefani

Wolfgang Wurst

Casper C Hoogenraad

Donald S Kirkpatrick

Damian Refojo

Morgan Sheng

Affiliations

Soon will be listed here.

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-εGG-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.

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