STK4 inhibits the E3 activity of HOIP by phosphorylating its allosteric ubiquitin-binding site.

IF 12.5 1区 生物学 Q1 CELL BIOLOGY
Yaru Wang, Xindi Zhou, Zhiqiao Lin, Yichao Huang, Yuchao Zhang, Haobo Liu, Yuqian Zhou, Jianping Liu, Lifeng Pan
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引用次数: 0

Abstract

HOIP, an RBR-type E3 ligase and the catalytic subunit of the linear ubiquitin chain assembly complex (LUBAC), plays crucial roles in various cellular processes, including the NF-κB signaling pathway. The E3 activity of HOIP can be inhibited by the kinase STK4-mediated phosphorylation, although the mechanism is poorly understood. In this study, using biochemical, mass spectrometry and structural approaches, we systemically characterize the association of STK4 with HOIP, and unveil that STK4 can directly bind to the RING2-LDD module of HOIP through its kinase domain. The determined crystal structure of STK4 in complex with HOIP RING2-LDD not only elucidates the detailed binding mechanism of STK4 with HOIP, but also uncovers, for the first time, a unique binding mode of STK4 with its substrate. Moreover, we reveal that STK4 can directly phosphorylate the T786 residue of HOIP that is located in the allosteric ubiquitin-binding site of HOIP. Importantly, the phosphorylation of HOIP T786 mediated by STK4 can block the binding of ubiquitin to the allosteric site of HOIP, thereby attenuating the E3 activity of HOIP. In all, our findings provide mechanistic insights into the interaction between STK4 and HOIP as well as the negative regulation of HOIP's E3 activity by STK4-mediated phosphorylation, which are valuable for further understanding the regulatory modes of RBR-type E3 ligases.

STK4通过磷酸化其变构泛素结合位点来抑制HOIP的E3活性。
HOIP是一种rbr型E3连接酶,是线性泛素链组装复合物(LUBAC)的催化亚基,在包括NF-κB信号通路在内的多种细胞过程中起着至关重要的作用。HOIP的E3活性可以被激酶stk4介导的磷酸化所抑制,尽管其机制尚不清楚。本研究采用生化、质谱和结构分析等方法,系统表征了STK4与HOIP的关联,揭示了STK4可以通过其激酶结构域直接结合到HOIP的RING2-LDD模块上。确定了STK4与HOIP复合物RING2-LDD的晶体结构,不仅阐明了STK4与HOIP结合的详细机制,而且首次揭示了STK4与其底物的独特结合模式。此外,我们发现STK4可以直接磷酸化HOIP中位于HOIP变构泛素结合位点的T786残基。重要的是,STK4介导的HOIP T786磷酸化可以阻断泛素与HOIP变构位点的结合,从而减弱HOIP的E3活性。总之,我们的研究结果提供了STK4与HOIP相互作用的机制见解,以及STK4介导的磷酸化对HOIP E3活性的负调控,这对进一步了解rbr型E3连接酶的调控模式有价值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cell Discovery
Cell Discovery Biochemistry, Genetics and Molecular Biology-Molecular Biology
CiteScore
24.20
自引率
0.60%
发文量
120
审稿时长
20 weeks
期刊介绍: Cell Discovery is a cutting-edge, open access journal published by Springer Nature in collaboration with the Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences (CAS). Our aim is to provide a dynamic and accessible platform for scientists to showcase their exceptional original research. Cell Discovery covers a wide range of topics within the fields of molecular and cell biology. We eagerly publish results of great significance and that are of broad interest to the scientific community. With an international authorship and a focus on basic life sciences, our journal is a valued member of Springer Nature's prestigious Molecular Cell Biology journals. In summary, Cell Discovery offers a fresh approach to scholarly publishing, enabling scientists from around the world to share their exceptional findings in molecular and cell biology.
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