Department of Microbiology, Yonsei University College of Medicine, 50-1 Yonsei-Ro, Seodaemun-Gu, Seoul, 03722, South Korea.
Brain Korea 21 FOUR Project for Medical Science, Yonsei University College of Medicine, Seoul, 03722, South Korea.
Mol Med. 2022 Dec 30;28(1):164. doi: 10.1186/s10020-022-00596-0.
High mobility group box 1 (HMGB1) is a damage-associated molecular pattern (DAMP) molecule that plays a central role in innate immunity. HMGB1 acts as a late mediator of inflammation when actively secreted in response to inflammatory stimuli. Several post-translational modifications (PTMs), including acetylation, phosphorylation, and oxidation, are involved in HMGB1 secretion. However, the E3 ligases of HMGB1 and the mechanism by which DUBs regulate HMGB1 deubiquitination are not well known.
LC-MS/MS, proximity ligation assay, immunoprecipitation were used to identify ubiquitin-specific protease 13 (USP13) as a binding partner of HMGB1 and to investigate ubiquitination of HMGB1. USP13 domain mutant was constructed for domain study and Spautin-1 was treated for inhibition of USP13. Confocal microscopy image showed localization of HMGB1 by USP13 overexpression. The data were analyzed using one-way analysis of variance with Tukey's honestly significant difference post-hoc test for multiple comparisons or a two-tailed Student's t-test.
We identified ubiquitin-specific protease 13 (USP13) as a novel binding partner of HMGB1 and demonstrated that USP13 plays a role in stabilizing HMGB1 from ubiquitin-mediated degradation. USP13 overexpression increased nucleocytoplasmic translocation of HMGB1 and promoted its secretion, which was inhibited by treatment with Spautin-1, a selective inhibitor of USP13.
Taken together, we suggest that USP13 is a novel deubiquitinase of HMGB1 that regulates the stability and secretion of HMGB1.
高迁移率族蛋白 B1（HMGB1）是一种损伤相关分子模式（DAMP）分子，在天然免疫中发挥核心作用。HMGB1 作为一种晚期炎症介质，在对炎症刺激的主动分泌时发挥作用。几种翻译后修饰（PTMs），包括乙酰化、磷酸化和氧化，参与 HMGB1 的分泌。然而，HMGB1 的 E3 连接酶和 DUBs 调节 HMGB1 去泛素化的机制尚不清楚。
使用 LC-MS/MS、邻近连接测定法和免疫沉淀法，鉴定出泛素特异性蛋白酶 13（USP13）是 HMGB1 的结合伴侣，并研究 HMGB1 的泛素化。构建 USP13 结构域突变体进行结构域研究，并用 Spautin-1 处理抑制 USP13。通过过表达 USP13 显示共聚焦显微镜图像中 HMGB1 的定位。使用单因素方差分析和 Tukey 事后检验进行多重比较或双尾学生 t 检验对数据进行分析。
我们鉴定出泛素特异性蛋白酶 13（USP13）是 HMGB1 的一种新型结合伴侣，并证明 USP13 在稳定 HMGB1 免受泛素介导的降解方面发挥作用。USP13 的过表达增加了 HMGB1 的核质易位，并促进其分泌，这一过程可被 USP13 的选择性抑制剂 Spautin-1 抑制。
综上所述，我们认为 USP13 是 HMGB1 的一种新型去泛素酶，调节 HMGB1 的稳定性和分泌。
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