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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Experimental interrogation of the path dependence and stochasticity of protein evolution using phage-assisted continuous evolution
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Experimental interrogation of the path dependence and stochasticity of protein evolution using phage-assisted continuous evolution

机译：使用噬菌体辅助的连续进化对蛋白质进化的路径依赖性和随机性进行实验性询问

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摘要

To what extent are evolutionary outcomes determined by a population's recent environment, and to what extent do they depend on historical contingency and random chance? Here we apply a unique experimental system to investigate evolutionary reproducibility and path dependence at the protein level. We combined phage-assisted continuous evolution with high-throughput sequencing to analyze evolving protein populations as they adapted to divergent and then convergent selection pressures over hundreds of generations. Independent populations of T7 RNA polymerase genes were subjected to one of two selection histories (“pathways”) demanding recognition of distinct intermediate promoters followed by a common final promoter. We observed distinct classes of solutions with unequal phenotypic activity and evolutionary potential evolve from the two pathways, as well as from replicate populations exposed to identical selection conditions. Mutational analysis revealed specific epistatic interactions that explained the observed path dependence and irreproducibility. Our results reveal in molecular detail how protein adaptation to different environments, as well as stochasticity among populations evolved in the same environment, can both generate evolutionary outcomes that preclude subsequent convergence.
机译：进化的结果在多大程度上取决于人口的近期环境，它们在多大程度上取决于历史偶然性和随机机会？在这里，我们应用独特的实验系统来研究蛋白质水平上的进化重现性和路径依赖性。我们将噬菌体辅助的连续进化与高通量测序相结合，以分析不断进化的蛋白质种群，因为它们适应了数百代人的发散然后收敛的选择压力。对T7 RNA聚合酶基因的独立种群进行两种选择历史（“途径”）之一，要求识别不同的中间启动子，然后再识别共同的最终启动子。我们观察到具有不同表型活性和进化潜力的不同类别的解决方案是从这两种途径以及暴露于相同选择条件的复制种群中进化而来的。突变分析揭示了特定的上位性相互作用，这解释了观察到的路径依赖性和不可再现性。我们的研究结果从分子细节上揭示了蛋白质对不同环境的适应性以及在同一环境中进化的种群之间的随机性如何都能产生进化结果，从而阻止随后的收敛。

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