Sears Rhiannon M, Nowling Nathan L, Yarbro Jake, Zhao Ning
Department of Biochemistry and Molecular Genetics, University of Colorado-Anschutz Medical Campus, Aurora, CO, U.S.A.
Biochem J. 2025 Jan 30;482(3):BCJ20240183. doi: 10.1042/BCJ20240183.
Translation is a highly regulated process that includes three steps: initiation, elongation, and termination. Tremendous efforts have been spent to study the regulation of each translation step. In the last two decades, researchers have begun to investigate translation by tracking it in its native and live intracellular environment with high spatiotemporal resolution. To achieve this goal, a handful of tagging tools have been developed that can distinguish nascent chains from previously synthesized mature proteins. In this review, we will focus on these tagging tools and describe their development, working mechanisms, and advantages and drawbacks in tracking translation in live mammalian cells and organisms. In the second part of the review, we will summarize novel discoveries in translation by a recently developed nascent polypeptide tracking technology using tandem epitope tag array tagging tools. The superior spatiotemporal resolution of this technology enables us to directly and continuously track nascent chains live and thus reveal preferred translation location and timing, as well as the kinetics of canonical and noncanonical translation, translation bursts, ribosome quality control, and nonsense-mediated mRNA decay. In the future, we expect more tagging tools to be developed that allow us to track other regulation processes of a protein, such as folding, modifications, and degradation. With the expanding tagging toolbox, there is potential that we can track a protein from translation to degradation to fully understand its regulation in a native live cell environment.
翻译是一个受到高度调控的过程，包括起始、延伸和终止三个步骤。人们已经付出了巨大努力来研究每个翻译步骤的调控。在过去二十年中，研究人员开始通过在其天然的活细胞内环境中以高时空分辨率追踪翻译过程来进行研究。为实现这一目标，已经开发了一些标记工具，这些工具可以区分新生链与先前合成的成熟蛋白质。在这篇综述中，我们将聚焦于这些标记工具，并描述它们在活的哺乳动物细胞和生物体中追踪翻译时的开发、工作机制以及优缺点。在综述的第二部分，我们将总结使用串联表位标签阵列标记工具的最新开发的新生多肽追踪技术在翻译方面的新发现。这项技术卓越的时空分辨率使我们能够直接且持续地实时追踪新生链，从而揭示翻译的优先位置和时间，以及经典和非经典翻译、翻译爆发、核糖体质量控制和无义介导的mRNA降解的动力学。未来，我们期望开发出更多的标记工具，使我们能够追踪蛋白质的其他调控过程，如折叠、修饰和降解。随着标记工具库的不断扩大，我们有可能追踪一种蛋白质从翻译到降解的全过程，以在天然的活细胞环境中充分理解其调控机制。
