Mapping the in vivo fitness landscape of a tethered ribosome.
Mapping the in vivo fitness landscape of a tethered ribosome.
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DOI：
10.1126/sciadv.ade8934
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发表时间：
2023-04-28
期刊：
影响因子：
13.6
通讯作者：
Isaacs, Farren J.
中科院分区：
文献类型：
--
作者：
Radford, Felix;Rinehart, Jesse;Isaacs, Farren J.
适合度景观是遗传元素序列空间的模型，它映射每个序列如何对应于其活动，并可用于指导实验室进化。核糖体是一种大分子机器，对所有生物体中的蛋白质合成至关重要。由于显性致死突变的流行，核糖体肽基转移中心（PTC）的全面的健身景观尚未实现。在这里，我们开发了一种方法来功能映射的正交拴系核糖体（oRiboT），它允许位于PTC和由此产生的上位相互作用的核苷酸的完全诱变。我们发现，大多数核苷酸研究显示出灵活的突变，并确定它们之间的上位相互作用，这补偿了有害的突变。这项工作为深入了解核糖体功能和延展性提供了基础，并可用于设计工程核糖体，并应用于合成下一代生物材料和治疗。核糖体的适应度景观是通过测定正交系留核糖体文库的翻译来计算的。
Fitness landscapes are models of the sequence space of a genetic element that map how each sequence corresponds to its activity and can be used to guide laboratory evolution. The ribosome is a macromolecular machine that is essential for protein synthesis in all organisms. Because of the prevalence of dominant lethal mutations, a comprehensive fitness landscape of the ribosomal peptidyl transfer center (PTC) has not yet been attained. Here, we develop a method to functionally map an orthogonal tethered ribosome (oRiboT), which permits complete mutagenesis of nucleotides located in the PTC and the resulting epistatic interactions. We found that most nucleotides studied showed flexibility to mutation, and identified epistatic interactions between them, which compensate for deleterious mutations. This work provides a basis for a deeper understanding of ribosome function and malleability and could be used to inform design of engineered ribosomes with applications to synthesize next-generation biomaterials and therapeutics. The fitness landscape of the ribosome is calculated by assaying translation by libraries of orthogonal tethered ribosomes.
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