Nature Chemical Biology ( IF 12.9 ) Pub Date : 2024-07-08 , DOI: 10.1038/s41589-024-01677-3 Gihoon Lee 1 , Tom W Muir 1
The post-translational regulation of protein function is involved in most cellular processes. As such, synthetic biology tools that operate at this level provide opportunities for manipulating cellular states. Here we deploy proximity-triggered protein trans-splicing technology to enable the time-resolved synthesis of target proteins from premade parts. The modularity of the strategy allows for the addition or removal of various control elements as a function of the splicing reaction, in the process permitting the cellular location and/or activity state of starting materials and products to be differentiated. The approach is applied to a diverse set of proteins, including the kinase oncofusions breakpoint cluster region–Abelson (BCR–ABL) and DNAJ–PKAc where dynamic cellular phosphorylation events are dissected, revealing distinct phases of signaling and identifying molecular players connecting the oncofusion to cancer transformation as new therapeutic targets of cancer cells. We envision that the tools and control strategies developed herein will allow the activity of both naturally occurring and designer proteins to be harnessed for basic and applied research.
中文翻译:
时间分辨蛋白质合成揭示细胞信号传导的不同阶段
蛋白质功能的翻译后调节参与大多数细胞过程。因此,在这个水平上运行的合成生物学工具为操纵细胞状态提供了机会。在这里,我们部署了邻近触发的蛋白质反式剪接技术,以实现从预制部件中时间分辨地合成目标蛋白质。该策略的模块化允许根据剪接反应添加或去除各种控制元件,在此过程中允许区分起始材料和产物的细胞位置和/或活性状态。该方法适用于多种蛋白质,包括激酶肿瘤融合断点簇区域 - Abelson (BCR-ABL) 和 DNAJ-PKAc,其中动态细胞磷酸化事件被剖析,揭示信号传导的不同阶段并识别将肿瘤融合与肿瘤融合相关的分子参与者。癌症转化作为癌细胞新的治疗靶点。我们设想,本文开发的工具和控制策略将允许利用天然存在的蛋白质和设计蛋白质的活性进行基础和应用研究。