CTLA4 的快速更替与可逆泛素化的复杂结构有关。

IF 7.4 1区 生物学 Q1 CELL BIOLOGY Journal of Cell Biology Pub Date : 2024-10-15 DOI:10.1083/jcb.202312141
Pei Yee Tey,Almut Dufner,Klaus-Peter Knobeloch,Jonathan N Pruneda,Michael J Clague,Sylvie Urbé
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引用次数: 0

摘要

免疫检查点调节因子 CTLA4 是一种异常短命的膜蛋白。在这里,我们发现它的溶酶体降解依赖于赖氨酸残基 203 和 213 的泛素化。在环己亚胺处理后,抑制 v-ATP 酶可部分恢复 CTLA4 的水平,但同时也发现了外泌体中分泌的部分 CTLA4。内体去泛素化酶 USP8 与 CTLA4 相互作用,失去 USP8 会增强 CTLA4 在癌细胞、小鼠 CD4+ T 细胞和癌细胞衍生的外泌体中的泛素化。USP8适配蛋白HD-PTP而非ESCRT-0的缺失再现了这种细胞表型,但在外泌体结合方面显示出不同的特性。重新表达野生型 USP8(但催化不活跃或定位受损的 ΔMIT 结构域突变体)不能挽救 CTLA4 的延迟降解,也不能抵消其在聚类内体中的积累。对与 CTLA4 相关的泛素链连接进行的 UbiCRest 分析发现,传统的 Lys63 与更不寻常的 Lys27 和 Lys29 连接的多泛素链形成了复杂的混合物,这可能是蛋白质快速周转的基础。
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Rapid turnover of CTLA4 is associated with a complex architecture of reversible ubiquitylation.
The immune checkpoint regulator CTLA4 is an unusually short-lived membrane protein. Here, we show that its lysosomal degradation is dependent on ubiquitylation at lysine residues 203 and 213. Inhibition of the v-ATPase partially restores CTLA4 levels following cycloheximide treatment, but also reveals a fraction that is secreted in exosomes. The endosomal deubiquitylase, USP8, interacts with CTLA4, and its loss enhances CTLA4 ubiquitylation in cancer cells, mouse CD4+ T cells, and cancer cell-derived exosomes. Depletion of the USP8 adapter protein, HD-PTP, but not ESCRT-0 recapitulates this cellular phenotype but shows distinct properties vis-à-vis exosome incorporation. Re-expression of wild-type USP8, but neither a catalytically inactive nor a localization-compromised ΔMIT domain mutant can rescue delayed degradation of CTLA4 or counteract its accumulation in clustered endosomes. UbiCRest analysis of CTLA4-associated ubiquitin chain linkages identifies a complex mixture of conventional Lys63- and more unusual Lys27- and Lys29-linked polyubiquitin chains that may underly the rapidity of protein turnover.
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来源期刊
Journal of Cell Biology
Journal of Cell Biology 生物-细胞生物学
CiteScore
12.60
自引率
2.60%
发文量
213
审稿时长
1 months
期刊介绍: The Journal of Cell Biology (JCB) is a comprehensive journal dedicated to publishing original discoveries across all realms of cell biology. We invite papers presenting novel cellular or molecular advancements in various domains of basic cell biology, along with applied cell biology research in diverse systems such as immunology, neurobiology, metabolism, virology, developmental biology, and plant biology. We enthusiastically welcome submissions showcasing significant findings of interest to cell biologists, irrespective of the experimental approach.
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