CRISPR筛选发现BAP1是调节SPIN4稳定性的去泛素酶。

IF 3 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry Biochemistry Pub Date : 2025-09-30 DOI:10.1021/acs.biochem.5c00493

Alondra Sanchez, Chen Zhou, Rima Tulaiha, Francisco Ramirez, Lu Wang, Xiaoyu Zhang

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引用次数: 0

摘要

蛋白质稳态是由E3泛素连接酶和去泛素酶（DUBs）的协同作用严格控制的。我们之前发现Spindlin-4 (SPIN4)，一种组蛋白H3K4me3读取器，是DCAF16的降解底物。在这项研究中，我们使用聚焦E3连接酶的CRISPR-Cas9敲除筛选证实了这种降解途径。此外，通过dub聚焦的CRISPR-Cas9敲除筛选和生化分析，我们证明了去泛素酶BAP1通过其去泛素化活性与SPIN4相互作用并稳定SPIN4。BAP1的抑制或缺失降低了SPIN4水平，突出了其在维持SPIN4稳态中的关键作用。蛋白质组学和相互作用组学分析进一步支持这一调控轴。这些发现揭示了控制SPIN4稳定性的动态平衡，对表观遗传调控和疾病过程具有潜在的意义。

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CRISPR Screen Identifies BAP1 as a Deubiquitinase Regulating SPIN4 Stability.

Protein homeostasis is tightly controlled by the coordinated actions of E3 ubiquitin ligases and deubiquitinases (DUBs). We previously identified Spindlin-4 (SPIN4), a histone H3K4me3 reader, as a degradation substrate of DCAF16. In this study, we confirmed this degradation pathway using an E3 ligase-focused CRISPR-Cas9 knockout screen. Furthermore, through a DUB-focused CRISPR-Cas9 knockout screen and biochemical analyses, we demonstrated that the deubiquitinase BAP1 interacts with and stabilizes SPIN4 via its deubiquitination activity. Inhibition or loss of BAP1 reduces SPIN4 levels, highlighting its critical role in maintaining SPIN4 homeostasis. Proteomics and interactome analyses further support this regulatory axis. These findings reveal a dynamic balance controlling SPIN4 stability, with potential implications for epigenetic regulation and disease processes.

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来源期刊

Biochemistry Biochemistry 生物-生化与分子生物学

CiteScore

5.50

自引率

3.40%

发文量

336

审稿时长

1-2 weeks

期刊介绍： Biochemistry provides an international forum for publishing exceptional, rigorous, high-impact research across all of biological chemistry. This broad scope includes studies on the chemical, physical, mechanistic, and/or structural basis of biological or cell function, and encompasses the fields of chemical biology, synthetic biology, disease biology, cell biology, nucleic acid biology, neuroscience, structural biology, and biophysics. In addition to traditional Research Articles, Biochemistry also publishes Communications, Viewpoints, and Perspectives, as well as From the Bench articles that report new methods of particular interest to the biological chemistry community.