Androniciuc Ana-Miruna, Tate Edward W, Vincent Jean-Paul
The Francis Crick Institute, London, England, United Kingdom.
Department of Chemistry, Imperial College London, London, England, United Kingdom.
MicroPubl Biol. 2024 May 29;2024. doi: 10.17912/micropub.biology.001210. eCollection 2024.
Wnt signalling coordinates growth and cell fate decisions during development and mis-regulation of Wnt signalling in adults is associated with a range of conditions, including cancer and neurodegenerative diseases. Therefore, means of modulating Wnt proteins and/or cofactors could have significant therapeutic potential. As a first step towards enumerating the Wnt interactome, we devised an proximity labelling strategy to identify proteins that interact with Wingless (Wg), the main Wnt. We engineered the locus to express a functional TurboID-Wg fusion at endogenous levels and identified interactors by streptavidin pull-down from embryos, followed by mass spectrometry. Further analysis may in future extend the screen coverage and deliver functional validation of the newly identified interactors.
Wnt信号通路在发育过程中协调生长和细胞命运决定，而在成体中Wnt信号通路的失调与一系列病症相关，包括癌症和神经退行性疾病。因此，调节Wnt蛋白和/或辅助因子的方法可能具有重大的治疗潜力。作为列举Wnt相互作用组的第一步，我们设计了一种邻近标记策略，以鉴定与主要Wnt——无翅型（Wg）相互作用的蛋白质。我们对该基因座进行工程改造，以在内源水平表达功能性TurboID-Wg融合蛋白，并通过从胚胎中进行链霉亲和素下拉，随后进行质谱分析来鉴定相互作用蛋白。未来的进一步分析可能会扩大筛选范围，并对新鉴定的相互作用蛋白进行功能验证。
