Synthetic Biology Pathway to Nucleoside Triphosphates for Expanded Genetic Alphabets.
Synthetic Biology Pathway to Nucleoside Triphosphates for Expanded Genetic Alphabets.
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扩展遗传字母表的核苷三磷酸的合成生物学途径。
DOI：
10.1021/acssynbio.3c00060
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发表时间：
2023
影响因子：
4.7
通讯作者：
Benner,StevenA
中科院分区：
文献类型：
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作者：
Li,Yubing;Abraham,Clay;Suslov,Oleg;Yaren,Ozlem;Shaw,RyanW;Kim,Myong-Jung;Wan,Shuo;Marliere,Philippe;Benner,StevenA
合成生物学的一个领域是寻找DNA的替代形式，以存储、转录和支持生物信息的进化。在这里，氢键供体和受体基团在沃森-克里克几何结构内重排以得到形成6个独立复制对的12个核苷酸。这种人工扩增的遗传信息系统（AEGIS）支持体外达尔文进化论。为了将AEGIS转移到活细胞中，接下来需要通过代谢途径从核苷中经济地制备AEGIS三磷酸盐，从而无需在生长培养基中添加这些昂贵的化合物。我们报告说，“多磷酸激酶”可以招募这样的途径，与天然二磷酸激酶和工程核苷激酶。这种途径在体外使AEGIS三磷酸盐，包括第三代三磷酸盐具有改善的在活细菌细胞中存活的能力。在这里首次生产的α-32 P标记形式中，它们被用于研究DNA聚合酶，发现第三代AEGIS三磷酸盐在天然酶中的表现优于第二代AEGIS三磷酸盐。
One horizon in synthetic biology seeks alternative forms of DNA that store, transcribe, and support the evolution of biological information. Here, hydrogen bond donor and acceptor groups are rearranged within a Watson–Crick geometry to get 12 nucleotides that form 6 independently replicating pairs. Such artificially expanded genetic information systems (AEGIS) support Darwinian evolutionin vitro. To move AEGIS into living cells, metabolic pathways are next required to make AEGIS triphosphates economically from their nucleosides, eliminating the need to feed these expensive compounds in growth media. We report that “polyphosphate kinases” can be recruited for such pathways, working with natural diphosphate kinases and engineered nucleoside kinases. This pathwayin vitromakes AEGIS triphosphates, including third-generation triphosphates having improved ability to survive in living bacterial cells. In α-32P-labeled forms, produced here for the first time, they were used to study DNA polymerases, finding cases where third-generation AEGIS triphosphates perform better with natural enzymes than second-generation AEGIS triphosphates.