Kyei-Baffour Edwin Sakyi, Bak Jitske, Silva Joana, Faller William J, Alkan Ferhat
Division of Oncogenomics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands.
Oncode Institute, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands.
NAR Genom Bioinform. 2025 May 8;7(2):lqaf045. doi: 10.1093/nargab/lqaf045. eCollection 2025 Jun.
It has become clear in recent years that ribosomes regularly stall during translation. Such translation impairment has many causes, including exposure to ribotoxic stress agents, the presence of specific RNA structures or sequences, or a shortage of amino acids or translation factors. If they are not resolved, stalled ribosomes can lead to ribosome collisions that are continuously surveilled by various sensor proteins. This in turn initiates a cascade of signalling events that can change the physiology and behaviour of cells. However, measuring changes in collision abundance has proved challenging, and as a result, the importance of collision-mediated biological responses is still unclear. Here, we show that computational analyses of standard ribosome profiling (Ribo-seq) data enable the prediction of changes in ribosome collisions between conditions. This is achieved by using the known 3D structure of collided ribosomes to define the ribosomal RNA (rRNA) positions that are differentially digested by RNases during the Ribo-seq protocol. Comparison of the relative rRNA reads at these positions allows the relative quantification of collisions between samples, an approach we call differential ribosome collisions by Analysis of rRNA Fragments (dricARF). When applied to public datasets across multiple organisms, our approach detects changes in collision events with unprecedented accuracy and sensitivity. In addition to providing supplementary evidence for ribosome collisions, our tool has the potential to uncover novel biological processes that are mediated by them. dricARF is available as part of the ARF R package and can be accessed through https://github.com/fallerlab/ARF.
近年来已经明确,核糖体在翻译过程中经常停滞。这种翻译障碍有多种原因,包括暴露于核糖体毒性应激剂、特定RNA结构或序列的存在,或氨基酸或翻译因子的短缺。如果这些问题得不到解决,停滞的核糖体可能会导致核糖体碰撞,而各种传感蛋白会持续监测这种碰撞。这进而引发一系列信号事件,可能会改变细胞的生理和行为。然而,事实证明,测量碰撞丰度的变化具有挑战性,因此,碰撞介导的生物学反应的重要性仍不清楚。在这里,我们表明,对标准核糖体分析(Ribo-seq)数据进行计算分析能够预测不同条件下核糖体碰撞的变化。这是通过利用碰撞核糖体的已知三维结构来定义在Ribo-seq实验过程中被核糖核酸酶差异消化的核糖体RNA(rRNA)位置来实现的。比较这些位置上的相对rRNA读数,可以对样品之间的碰撞进行相对定量,我们将这种方法称为通过rRNA片段分析进行差异核糖体碰撞(dricARF)。当应用于多种生物的公共数据集时,我们的方法以前所未有的准确性和灵敏度检测碰撞事件的变化。除了为核糖体碰撞提供补充证据外,我们的工具还有可能揭示由它们介导的新的生物学过程。dricARF作为ARF R包的一部分可用,可通过https://github.com/fallerlab/ARF访问。