Frank Daniel, Garnish Sarah E, Sandow Jarrod J, Weir Ashley, Liu Lin, Clayer Elise, Meza Lizeth, Rashidi Maryam, Cobbold Simon A, Scutts Simon R, Doerflinger Marcel, Anderton Holly, Lawlor Kate E, Lalaoui Najoua, Kueh Andrew J, Eng Vik Ven, Ambrose Rebecca L, Herold Marco J, Samson Andre L, Feltham Rebecca, Murphy James M, Ebert Gregor, Pearson Jaclyn S, Vince James E
The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Melbourne, VIC 3052, Australia.
Department of Medical Biology, The University of Melbourne, Parkville, VIC 3050, Australia.
iScience. 2022 Jun 17;25(7):104632. doi: 10.1016/j.isci.2022.104632. eCollection 2022 Jul 15.
Pathogen recognition and TNF receptors signal via receptor interacting serine/threonine kinase-3 (RIPK3) to cause cell death, including MLKL-mediated necroptosis and caspase-8-dependent apoptosis. However, the post-translational control of RIPK3 is not fully understood. Using mass-spectrometry, we identified that RIPK3 is ubiquitylated on K469. The expression of mutant RIPK3 K469R demonstrated that RIPK3 ubiquitylation can limit both RIPK3-mediated apoptosis and necroptosis. The enhanced cell death of overexpressed RIPK3 K469R and activated endogenous RIPK3 correlated with an overall increase in RIPK3 ubiquitylation. mice challenged with displayed enhanced bacterial loads and reduced serum IFNγ. However, macrophages and dermal fibroblasts were not sensitized to RIPK3-mediated apoptotic or necroptotic signaling suggesting that, in these cells, there is functional redundancy with alternate RIPK3 ubiquitin-modified sites. Consistent with this idea, the mutation of other ubiquitylated RIPK3 residues also increased RIPK3 hyper-ubiquitylation and cell death. Therefore, the targeted ubiquitylation of RIPK3 may act as either a brake or accelerator of RIPK3-dependent killing.
病原体识别和肿瘤坏死因子受体通过受体相互作用丝氨酸/苏氨酸激酶-3（RIPK3）发出信号，从而导致细胞死亡，包括混合谱系激酶结构域样蛋白（MLKL）介导的坏死性凋亡和半胱天冬酶-8依赖性凋亡。然而，RIPK3的翻译后调控尚未完全明确。通过质谱分析，我们鉴定出RIPK3在K469位点发生泛素化。突变型RIPK3 K469R的表达表明，RIPK3泛素化可限制RIPK3介导的凋亡和坏死性凋亡。过表达的RIPK3 K469R和活化的内源性RIPK3增强的细胞死亡与RIPK3泛素化的总体增加相关。用……攻击的小鼠表现出细菌载量增加和血清干扰素γ降低。然而，……巨噬细胞和真皮成纤维细胞对RIPK3介导的凋亡或坏死性凋亡信号不敏感，这表明在这些细胞中，存在与其他RIPK3泛素修饰位点的功能冗余。与此观点一致，其他泛素化的RIPK3残基的突变也增加了RIPK3的过度泛素化和细胞死亡。因此，RIPK3的靶向泛素化可能作为RIPK3依赖性杀伤的制动器或加速器。
