Emergence of ATP- and GTP-Binding Aptamers from Single RNA Sequences by Error-Prone Replication and Selection**

IF 3.1 Q2 CHEMISTRY, MULTIDISCIPLINARY ChemSystemsChem Pub Date : 2023-06-05 DOI:10.1002/syst.202300006
Dr. Falk Wachowius, Dr. Benjamin T. Porebski, Dr. Christopher M. Johnson, Dr. Philipp Holliger
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Abstract

The spontaneous emergence of function from diverse RNA sequence pools is widely considered an important transition in the origin of life. Here we show that diverse sequence pools are not a prerequisite for the emergence of function. Starting five independent selection experiments each from a single RNA seed sequence - comprising a central homopolymeric poly-A (or poly-U) segment flanked by different conserved primer binding sites - we observe transformation (continuous drift) of the seeds into low diversity sequence pools by mutation, truncation and recombination without ever reaching that of a random pool even after 24 rounds. Upon continuous error prone replication and selection for ATP binding we isolate specific ATP- or GTP-binding aptamers with low micromolar affinities. Our results have implications for early RNA evolution in the light of the high mutation rates associated with both non-enzymatic and enzymatic prebiotic RNA replication.

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通过易错复制和选择从单个RNA序列中产生ATP和GTP结合适体
从不同的RNA序列池中自发产生的功能被广泛认为是生命起源的一个重要转变。在这里,我们表明多样化的序列池并不是功能出现的先决条件。我们从单个RNA种子序列开始了五个独立的选择实验,每个选择实验都包括一个中心的同源聚a(或聚u)片段,两侧是不同的保守引物结合位点。我们观察到种子通过突变、截断和重组转化(连续漂移)到低多样性序列库,即使在24轮之后也没有达到随机序列库的水平。经过连续的错误复制和ATP结合的选择,我们分离出具有低微摩尔亲和力的特异性ATP或gtp结合适体。我们的研究结果对早期RNA进化具有启示意义,因为与非酶和酶促益生元RNA复制相关的高突变率。
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