MONITTR 可对秀丽隐杆线虫的转录和内源蛋白进行实时成像。

IF 7.4 1区 生物学 Q1 CELL BIOLOGY Journal of Cell Biology Pub Date : 2024-10-14 DOI:10.1083/jcb.202403198
Xiaofan Liu,Zhi Chang,Pingping Sun,Beibei Cao,Yuzhi Wang,Jie Fang,Yechun Pei,Baohui Chen,Wei Zou
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引用次数: 0

摘要

要在压力下最大限度地提高细胞存活率,就必须快速、瞬时地调整 RNA 和蛋白质的合成。然而,在单细胞水平和整个生物体内捕捉这些动态变化一直是个挑战。在这里,我们开发了一种名为 MONITTR(MS2-嵌入式 mCherry 转录监测)的系统,用于实时同步测量秀丽隐杆线虫的新生转录本和内源蛋白质水平。利用该系统,我们监测了禁食诱导基因的转录突变,发现表皮通过调节活跃转录核的比例和单个等位基因的转录动力学对禁食做出反应。此外,我们的研究结果还揭示了转录因子NHR-49和HLH-30在管理禁食诱导基因转录动力学中的重要作用。此外,我们还追踪了热休克反应和ER未折叠蛋白反应期间的转录动态,观察到新生转录本水平在应激条件下的快速变化。总之,我们的研究为定量研究动物在各种生理和病理条件下如何时空调节转录提供了基础。
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MONITTR allows real-time imaging of transcription and endogenous proteins in C. elegans.
Maximizing cell survival under stress requires rapid and transient adjustments of RNA and protein synthesis. However, capturing these dynamic changes at both single-cell level and across an organism has been challenging. Here, we developed a system named MONITTR (MS2-embedded mCherry-based monitoring of transcription) for real-time simultaneous measurement of nascent transcripts and endogenous protein levels in C. elegans. Utilizing this system, we monitored the transcriptional bursting of fasting-induced genes and found that the epidermis responds to fasting by modulating the proportion of actively transcribing nuclei and transcriptional kinetics of individual alleles. Additionally, our findings revealed the essential roles of the transcription factors NHR-49 and HLH-30 in governing the transcriptional kinetics of fasting-induced genes under fasting. Furthermore, we tracked transcriptional dynamics during heat-shock response and ER unfolded protein response and observed rapid changes in the level of nascent transcripts under stress conditions. Collectively, our study provides a foundation for quantitatively investigating how animals spatiotemporally modulate transcription in various physiological and pathological conditions.
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来源期刊
Journal of Cell Biology
Journal of Cell Biology 生物-细胞生物学
CiteScore
12.60
自引率
2.60%
发文量
213
审稿时长
1 months
期刊介绍: The Journal of Cell Biology (JCB) is a comprehensive journal dedicated to publishing original discoveries across all realms of cell biology. We invite papers presenting novel cellular or molecular advancements in various domains of basic cell biology, along with applied cell biology research in diverse systems such as immunology, neurobiology, metabolism, virology, developmental biology, and plant biology. We enthusiastically welcome submissions showcasing significant findings of interest to cell biologists, irrespective of the experimental approach.
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