小柯机器人

单细胞Ribo-seq揭示细胞周期依赖性的翻译停顿
2021-09-11 18:49

近日,荷兰乌得勒支大学医学中心Alexander van Oudenaarden、Michael VanInsberghe等研究人员合作开发出单细胞Ribo-seq技术,揭示细胞周期依赖性的翻译停顿。2021年9月8日,《自然》杂志在线发表了这项成果。

据研究人员介绍,单细胞测序方法使人们能够深入分析各种生物体内细胞类型和细胞状态的多样性。这些工具主要侧重于对单细胞的基因组、表观基因组和转录组进行测序。然而,尽管最近在以单细胞分辨率通过质谱检测蛋白质方面取得了进展,但测量单个细胞的翻译仍然是一个重大挑战。

在现有方案的基础上,研究人员大幅提高了这些检测的灵敏度,以便在单细胞中进行核糖体分析。与机器学习方法相结合,这项技术实现了单密码子分辨率。研究人员通过证明对某一特定氨基酸的限制会导致核糖体在编码该氨基酸密码子的一个亚群上停顿来验证了这一方法。

值得注意的是,这种暂停只在一个与细胞周期状态相关的细胞亚群中观察到。研究人员在非限制性条件下进一步扩展了这一现象,并在有丝分裂期间检测到明显的GAA停顿。最后,研究人员证明了这项技术对罕见原发性肠内分泌细胞的适用性。这项技术为确定翻译过程对相似细胞之间显著多样性的贡献迈出了第一步。

附:英文原文

Title: Single-cell Ribo-seq reveals cell cycle-dependent translational pausing

Author: VanInsberghe, Michael, van den Berg, Jeroen, Andersson-Rolf, Amanda, Clevers, Hans, van Oudenaarden, Alexander

Issue&Volume: 2021-09-08

Abstract: Single-cell sequencing methods have enabled in-depth analysis of the diversity of cell types and cell states in a wide range of organisms. These tools focus predominantly on sequencing the genomes1, epigenomes2 and transcriptomes3 of single cells. However, despite recent progress in detecting proteins by mass spectrometry with single-cell resolution4, it remains a major challenge to measure translation in individual cells. Here, building on existing protocols5,6,7, we have substantially increased the sensitivity of these assays to enable ribosome profiling in single cells. Integrated with a machine learning approach, this technology achieves single-codon resolution. We validate this method by demonstrating that limitation for a particular amino acid causes ribosome pausing at a subset of the codons encoding the amino acid. Of note, this pausing is only observed in a sub-population of cells correlating to its cell cycle state. We further expand on this phenomenon in non-limiting conditions and detect pronounced GAA pausing during mitosis. Finally, we demonstrate the applicability of this technique to rare primary enteroendocrine cells. This technology provides a first step towards determining the contribution of the translational process to the remarkable diversity between seemingly identical cells.

DOI: 10.1038/s41586-021-03887-4

Source: https://www.nature.com/articles/s41586-021-03887-4

Nature:《自然》,创刊于1869年。隶属于施普林格·自然出版集团,最新IF:69.504
官方网址:http://www.nature.com/
投稿链接:http://www.nature.com/authors/submit_manuscript.html


本期文章:《自然》:Online/在线发表

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