Hernández-Quiles Miguel, Baak Rosalie, Borgman Anouska, den Haan Suzanne, Sobrevals Alcaraz Paula, van Es Robert, Kiss-Toth Endre, Vos Harmjan, Kalkhoven Eric
Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, 3584 CG Utrecht, The Netherlands.
Oncode Institute and Molecular Cancer Research, Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, 3584 CG Utrecht, The Netherlands.
Cancers (Basel). 2021 Dec 16;13(24):6318. doi: 10.3390/cancers13246318.
The three human Tribbles (TRIB) pseudokinases have been implicated in a plethora of signaling and metabolic processes linked to cancer initiation and progression and can potentially be used as biomarkers of disease and prognosis. While their modes of action reported so far center around protein-protein interactions, the comprehensive profiling of TRIB interactomes has not been reported yet. Here, we have developed a robust mass spectrometry (MS)-based proteomics approach to characterize Tribbles' interactomes and report a comprehensive assessment and comparison of the TRIB1, -2 and -3 interactomes, as well as domain-specific interactions for TRIB3. Interestingly, TRIB3, which is predominantly localized in the nucleus, interacts with multiple transcriptional regulators, including proteins involved in gene repression. Indeed, we found that TRIB3 repressed gene transcription when tethered to DNA in breast cancer cells. Taken together, our comprehensive proteomic assessment reveals previously unknown interacting partners and functions of Tribbles proteins that expand our understanding of this family of proteins. In addition, our findings show that MS-based proteomics provides a powerful tool to unravel novel pseudokinase biology.
三种人类 Tribbles（TRIB）假激酶与大量与癌症发生和进展相关的信号传导和代谢过程有关，并且有可能用作疾病和预后的生物标志物。虽然目前报道的它们的作用方式主要围绕蛋白质 - 蛋白质相互作用，但尚未有关于 TRIB 相互作用组的全面分析报道。在此，我们开发了一种基于质谱（MS）的强大蛋白质组学方法来表征 Tribbles 的相互作用组，并报告了对 TRIB1、-2 和 -3 相互作用组的全面评估和比较，以及 TRIB3 的结构域特异性相互作用。有趣的是，主要定位于细胞核的 TRIB3 与多种转录调节因子相互作用，包括参与基因抑制的蛋白质。事实上，我们发现当在乳腺癌细胞中与 DNA 结合时，TRIB3 会抑制基因转录。综上所述，我们的全面蛋白质组学评估揭示了 Tribbles 蛋白以前未知的相互作用伙伴和功能，扩展了我们对这一蛋白质家族的理解。此外，我们的研究结果表明基于 MS 的蛋白质组学为揭示新的假激酶生物学提供了一个强大的工具。
