使用外部标准对急性压力诱导的核糖体占用水平变化进行转录组范围的分析
Annie W Shieh1, Sandeep K Bansal1, Zhen Zuo2
1Center for Human Genetics, The Brown foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX, United States of America.
PloS one
|November 21, 2023
概括
新的spike-in寡合体准确地描述了细胞应激期间的mRNA翻译变化. 这些标准提高了核糖体分析的准确性,克服了压力反应研究传统规范化方法的局限性.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 细胞应激反应的应激反应
背景情况:
- 急性细胞应激会通过依赖上限的途径减少mRNA翻译.
- 选择性翻译对于细胞应激反应调节至关重要.
- 在压力下精确的转录组范围的翻译概况是具有挑战性的.
研究的目的:
- 开发和验证准确的核糖体造型的外部标准.
- 评估压力诱导的翻译变化的数据规范化方法.
- 为了解决压力研究中标准规范化技术引入的偏见.
主要方法:
- 设计并生产了一个由16个寡合体组成的面板,作为外部标准.
- 在酸诱导的氧化应激模型中利用了尖端的寡合物.
- 将TMM正常化与RUVg正常化使用尖端控制器进行比较.
主要成果:
- 尖端的寡合体显示了观察到的和预期的量化之间强烈的线性相关性.
- TMM正常化产生了87.5%的假阳性和3.25倍的转移.
- RUVg正常化准确地回顾了压力诱导的翻译减少,偏差最小.
结论:
- 开发的尖端寡合体是有效的外部标准,用于核糖体分析.
- 像TMM这样的全球缩放规范化方法不适合压力诱导的翻译研究.
- 使用尖端控制的RUVg规范化,在细胞应激期间提供准确和公正的翻译概况.
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