The discovery of extremophiles helped enable the development of groundbreaking technology such as polymerase chain reaction. Temperature variation is often an essential step of these technology platforms, but the effect of temperature on the error rate of polymerases from different origins is under-explored. Here, we applied high-throughput sequencing to profile the error rates of DNA polymerases from psychrophilic, mesophilic, and thermophilic origins with single-molecule resolution. We found that reaction temperature substantially increases substitution and deletion error rates of psychrophilic and mesophilic DNA polymerases. Our motif analysis shows that the substitution error profiles cluster according to phylogenetic similarity of polymerases, not reaction temperature, thus suggesting that reaction temperature increases global error rate of polymerases independent of sequence context. Intriguingly, we also found that the DNA polymerase I of a psychrophilic bacteria exhibits higher polymerization activity than its mesophilic ortholog across all temperature ranges, including down to −19°C which is well below the freezing temperature of water. Our results provide a useful reference for how reaction temperature, a crucial parameter of biochemistry, can affect DNA polymerase fidelity in organisms adapted to a wide range of thermal environments.

中文翻译：

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温度对聚合酶保真度的影响

极端微生物的发现有助于开发突破性技术，例如聚合酶链反应。温度变化通常是这些技术平台的重要步骤，但温度对不同来源聚合酶错误率的影响尚未得到充分探索。在这里，我们应用高通量测序以单分子分辨率分析来自嗜冷、嗜温和嗜热来源的 DNA 聚合酶的错误率。我们发现反应温度显着增加了嗜冷和嗜温 DNA 聚合酶的取代和缺失错误率。我们的基序分析表明，替换错误谱根据聚合酶的系统发育相似性而不是反应温度进行聚类，因此表明反应温度会增加聚合酶的全局错误率，而与序列背景无关。有趣的是，我们还发现嗜冷细菌的 DNA 聚合酶 I 在所有温度范围内都表现出比其嗜温直向同源物更高的聚合活性，包括低至 -19°C，这远低于水的冰点温度。我们的结果为反应温度（生物化学的一个关键参数）如何影响适应各种热环境的生物体的 DNA 聚合酶保真度提供了有用的参考。包括低至 -19°C，远低于水的冰点温度。我们的结果为反应温度（生物化学的一个关键参数）如何影响适应各种热环境的生物体的 DNA 聚合酶保真度提供了有用的参考。包括低至 -19°C，远低于水的冰点温度。我们的结果为反应温度（生物化学的一个关键参数）如何影响适应各种热环境的生物体的 DNA 聚合酶保真度提供了有用的参考。