Department of Biochemistry & Molecular Biology, University of Iowa, Iowa City, IA, USA.
Nucleic Acids Res. 2023 Sep 22;51(17):e92. doi: 10.1093/nar/gkad682.
Stochastic simulation models have played an important role in efforts to understand the mechanistic basis of prokaryotic transcription and translation. Despite the fundamental linkage of these processes in bacterial cells, however, most simulation models have been limited to representations of either transcription or translation. In addition, the available simulation models typically either attempt to recapitulate data from single-molecule experiments without considering cellular-scale high-throughput sequencing data or, conversely, seek to reproduce cellular-scale data without paying close attention to many of the mechanistic details. To address these limitations, we here present spotter (Simulation of Prokaryotic Operon Transcription & Translation Elongation Reactions), a flexible, user-friendly simulation model that offers highly-detailed combined representations of prokaryotic transcription, translation, and DNA supercoiling. In incorporating nascent transcript and ribosomal profiling sequencing data, spotter provides a critical bridge between data collected in single-molecule experiments and data collected at the cellular scale. Importantly, in addition to rapidly generating output that can be aggregated for comparison with next-generation sequencing and proteomics data, spotter produces residue-level positional information that can be used to visualize individual simulation trajectories in detail. We anticipate that spotter will be a useful tool in exploring the interplay of processes that are crucially linked in prokaryotes.
随机模拟模型在理解原核转录和翻译的机制基础方面发挥了重要作用。然而,尽管这些过程在细菌细胞中有着根本的联系,但大多数模拟模型仅限于转录或翻译的表示。此外,现有的模拟模型通常要么试图重现单分子实验的数据,而不考虑细胞尺度的高通量测序数据,要么相反,试图复制细胞尺度的数据,而不密切关注许多机制细节。为了解决这些限制,我们在这里提出了 spotter(原核操纵子转录和翻译延伸反应的模拟),这是一个灵活、用户友好的模拟模型,提供了原核转录、翻译和 DNA 超螺旋的高度详细的组合表示。在整合新生转录本和核糖体分析测序数据方面,spotter 在单细胞实验中收集的数据和在细胞尺度上收集的数据之间提供了一个关键的桥梁。重要的是,除了快速生成可用于与下一代测序和蛋白质组学数据进行比较的聚合输出外,spotter 还生成了残基级别的位置信息,可用于详细可视化单个模拟轨迹。我们预计 spotter 将成为探索在原核生物中紧密相关的过程相互作用的有用工具。