CRBM, University of Montpellier, CNRS, Montpellier, France.
Institute of Molecular Biology (IMB), Mainz, Germany.
Cell Rep. 2021 Jan 19;34(3):108635. doi: 10.1016/j.celrep.2020.108635.
The ubiquitin-like molecule NEDD8 controls several biological processes and is a promising target for therapeutic intervention. NEDDylation occurs through specific NEDD8 enzymes (canonical) or enzymes of the ubiquitin system (atypical). Identification of NEDD8 sites on substrates is critical for delineating the processes controlled by NEDDylation. By combining the use of the NEDD8 R74K mutant with anti-di-glycine (anti-diGly) antibodies, we identified 1,101 unique NEDDylation sites in 620 proteins. Bioinformatics analysis reveals that canonical and atypical NEDDylation have distinct proteomes; the spliceosome/mRNA surveillance/DNA replication and ribosome/proteasome, respectively. The data also reveal the formation of poly-NEDD8, hybrid NEDD8-ubiquitin, and NEDD8-SUMO-2 chains as potential molecular signals. In particular, NEDD8-SUMO-2 chains are induced upon proteotoxic stress (atypical) through NEDDylation of K11 in SUMO-2, and conjugates accumulate in previously described nucleolus-related inclusions. The study uncovers a diverse proteome for NEDDylation and is consistent with the concept of extensive cross-talk between ubiquitin and Ubls under proteotoxic stress conditions.
类泛素分子 NEDD8 调控多种生物学过程，是治疗干预的一个有前途的靶点。NEDDylation 通过特定的 NEDD8 酶（经典途径）或泛素系统的酶（非经典途径）发生。确定底物上的 NEDD8 位点对于描绘 NEDDylation 调控的过程至关重要。通过结合使用 NEDD8 R74K 突变体和抗二甘氨酸（anti-diGly）抗体，我们在 620 种蛋白质中鉴定出 1101 个独特的 NEDDylation 位点。生物信息学分析表明，经典和非经典 NEDDylation 具有不同的蛋白质组；剪接体/mRNA 监控/DNA 复制和核糖体/蛋白酶体。数据还揭示了多聚 NEDD8、杂交 NEDD8-泛素和 NEDD8-SUMO-2 链的形成作为潜在的分子信号。特别是，通过 SUMO-2 中 K11 的 NEDDylation，在蛋白毒性应激（非经典途径）下会诱导形成 NEDD8-SUMO-2 链，并且在先前描述的核仁相关包涵体中积累缀合物。该研究揭示了 NEDDylation 的广泛蛋白质组，并且与泛素和 Ubls 在蛋白毒性应激条件下广泛相互作用的概念一致。
