重塑细胞应激反应,以增强哺乳动物细胞中的遗传密码扩展
Mikhail E Sushkin1,2, Christine Koehler1,3, Edward A Lemke4,5
1Biocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128, Mainz, Germany.
Nature communications
|October 31, 2023
概括
哺乳动物细胞中的遗传密码扩展 (GCE) 效率通过对eIF2α通路进行工程改进而得到改善. 策略,包括添加PKR片段,提高翻译率和增强ncAA纳入蛋白质合成.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 遗传密码扩展 (GCE) 能够将非正规氨基酸 (ncAAs) 具体地纳入蛋白质中.
- 细胞应激反应,特别是PKR依赖的eIF2α酸化通路,可以通过降低哺乳动物细胞的翻译速率来限制GCE的效率.
研究的目的:
- 研究设计eIF2α通路的策略,以提高翻译率.
- 在哺乳动物细胞和设计器官中提高GCE的效率.
主要方法:
- 测试各种策略来设计eIF2α通路.
- 评估这些策略对翻译率和GCE效率的影响.
- 使用直角翻译设计器官 (OTOs) 进行mRNA选择性ncAA整合.
主要成果:
- 设计eIF2α路径显著提高了翻译速率.
- 干预措施,特别是N端PKR片段 (1-174) 的添加,可以显著增强细胞质GCE.
- 此外,PKR片段还可以提高OTO内部GCE的效率.
结论:
- 设计eIF2α通路的策略可以克服GCE效率的局限性.
- N-终端PKR片段是增强GCE在细胞质系统和设计器官中的一种强有力的工具.
- 这项工作提供了一种优化蛋白质翻译的方法,并扩大设计器官的功能.
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