Gutierrez Isaura Vanessa, Park Moonhee, Sar Lauren, Rodriguez Ryan, Snider Daltry L, Torres Gabriella, Scaglione K Matthew, Horner Stacy M
bioRxiv. 2025 Mar 19:2025.03.19.644084. doi: 10.1101/2025.03.19.644084.
Post-translational modifications are critical for regulating the RIG-I signaling pathway. Previously, we identified a role for the post-translation modification UFM1 (UFMylation) in promoting RIG-I signaling by stimulating the interaction between RIG-I and its membrane-targeting protein 14-3-3ε. Here, we identify UFMylation of 14-3-3ε as a novel regulatory mechanism promoting RIG-I signaling. We demonstrate that UFM1 conjugation to lysine residue K50 or K215 results in mono-UFMylation on 14-3-3ε and enhances its ability to promote RIG-I signaling. Importantly, we show that mutation of these residues (K50R/K215R) abolishes UFMylation and impairs induction of type I and III interferons without disrupting the interaction between 14-3-3ε and RIG-I. This suggests that UFMylation of 14-3-3ε likely stabilizes signaling events downstream of RIG-I activation to promote induction of interferon. Collectively, our work suggests that UFMylation-driven activation of 14-3-3ε facilitates innate immune signaling and highlights the broader role of UFMylation for antiviral defense and immune regulation.
Post-translational modifications provide regulatory control of antiviral innate immune responses. Our study reveals that UFMylation of 14-3-3ε is a control point for RIG-I-mediated antiviral signaling. We demonstrate that conjugation of UFM1 to specific lysine residues on 14-3-3ε enhances downstream signaling events that facilitate interferon induction, but surprisingly it does not affect 14-3-3ε binding to RIG-I. By identifying the precise sites of UFMylation on 14-3-3ε and their functional consequences, we provide insights into the regulatory layers governing antiviral innate immunity. These findings complement emerging evidence that UFMylation serves as a versatile modulator across diverse immune pathways. Furthermore, our work highlights how protein chaperones like 14-3-3ε can be dynamically modified to orchestrate complex signaling cascades, suggesting potential therapeutic approaches for targeting dysregulated innate immunity.
翻译后修饰对于调节RIG-I信号通路至关重要。此前，我们确定了翻译后修饰UFM1（UFMylation）通过刺激RIG-I与其膜靶向蛋白14-3-3ε之间的相互作用在促进RIG-I信号传导中的作用。在此，我们确定14-3-3ε的UFMylation是促进RIG-I信号传导的一种新型调节机制。我们证明，UFM1与赖氨酸残基K50或K215缀合会导致14-3-3ε上的单UFMylation，并增强其促进RIG-I信号传导的能力。重要的是，我们表明这些残基（K50R/K215R）的突变消除了UFMylation，并损害了I型和III型干扰素的诱导，而不会破坏14-3-3ε与RIG-I之间的相互作用。这表明14-3-3ε的UFMylation可能稳定RIG-I激活下游的信号事件，以促进干扰素的诱导。总体而言，我们的工作表明UFMylation驱动的14-3-3ε激活促进了先天免疫信号传导，并突出了UFMylation在抗病毒防御和免疫调节中的更广泛作用。
翻译后修饰为抗病毒先天免疫反应提供调节控制。我们的研究表明，14-3-3ε的UFMylation是RIG-I介导的抗病毒信号传导的控制点。我们证明，UFM1与14-3-3ε上特定赖氨酸残基的缀合增强了促进干扰素诱导的下游信号事件，但令人惊讶的是，它不影响14-3-3ε与RIG-I的结合。通过确定14-3-3ε上UFMylation的精确位点及其功能后果，我们深入了解了控制抗病毒先天免疫的调节层次。这些发现补充了新出现的证据，即UFMylation作为一种多功能调节剂存在于多种免疫途径中。此外，我们的工作突出了像14-3-3ε这样的蛋白质伴侣如何被动态修饰以协调复杂的信号级联反应，暗示了针对失调的先天免疫的潜在治疗方法。
