Data_Sheet_1_Neddylation Facilitates the Antiviral Response in Zebrafish.pdf

Neddylation is a type of post-translational protein modifications, in which neural precursor cell expressed developmentally downregulated protein 8 (NEDD8) is covalently conjugated to the lysine residues of target substrates. The best characterized principal substrates of neddylation are the cullin-RING ligases (CRLs). In addition, neddylation also modifies non-cullin proteins to affect gene regulation, cell survival, organ development, and stress response. However, the role of neddylation in antiviral innate immunity remain largely unknown. Here, we found that when neddylation was blocked by the NEDD8 activating enzyme E1 (NAE) inhibitor, MLN4924, the cellular and organismal antiviral response was suppressed. Moreover, the disruption of nedd8 increased the sensitivity of zebrafish to SVCV infection. Further assays indicated that blocking or silencing neddylation significantly downregulated key antiviral genes after poly (I:C) stimulation or SVCV infection, but dramatically increased SVCV replication. Neddylation of Irf3 and Irf7 was readily detected, but not of Mda5, Mavs, and Tbk1. Thus, our results not only demonstrated that neddylation facilitated the antiviral response in vitro and in vivo, but also revealed a novel role of nedd8 in antiviral innate immunity.

去乙酰化是一种翻译后蛋白质修饰类型，其中神经祖细胞表达的开发性下调蛋白8（NEDD8）通过共价键与靶底物中的赖氨酸残基结合。去乙酰化最为明确的主体底物为Cullin-RING连接酶（CRLs）。此外，去乙酰化亦能修饰非Cullin蛋白，进而影响基因调控、细胞存活、器官发育及应激反应。然而，去乙酰化在抗病毒先天免疫中的作用尚不明确。本研究发现，当NEDD8激活酶E1（NAE）抑制剂MLN4924阻断去乙酰化时，细胞和机体的抗病毒反应受到抑制。此外，NEDD8的破坏增加了斑马鱼对SVCV感染的敏感性。进一步的实验表明，阻断或沉默去乙酰化在poly(I:C)刺激或SVCV感染后显著下调了关键抗病毒基因，但显著增加了SVCV的复制。Irf3和Irf7的去乙酰化易于检测，而Mda5、Mavs和Tbk1则不易。因此，我们的研究结果不仅证明了去乙酰化在体外和体内促进了抗病毒反应，而且揭示了NEDD8在抗病毒先天免疫中的新型作用。