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MONITTR allows real-time imaging of transcription and endogenous proteins in C. elegans.
Journal of Cell Biology ( IF 7.4 ) 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

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.

中文翻译:


MONITTR 允许对秀丽隐杆线虫中的转录和内源性蛋白质进行实时成像。



在应激下最大限度地提高细胞存活率需要对 RNA 和蛋白质合成进行快速和瞬时调整。然而,在单细胞水平和整个生物体中捕获这些动态变化一直具有挑战性。在这里,我们开发了一个名为 MONITTR (MS2 嵌入的基于 mCherry 的转录监测)的系统,用于实时同时测量秀丽隐杆线虫中的新生转录本和内源性蛋白质水平。利用该系统,我们监测了禁食诱导基因的转录爆发,发现表皮通过调节主动转录细胞核的比例和单个等位基因的转录动力学来响应禁食。此外,我们的研究结果揭示了转录因子 NHR-49 和 HLH-30 在控制禁食下空腹诱导基因的转录动力学中的重要作用。此外,我们跟踪了热休克反应和 ER 未折叠蛋白反应期间的转录动力学,并观察到在应激条件下新生转录本水平的快速变化。总的来说,我们的研究为定量研究动物如何在各种生理和病理条件下时空调节转录提供了基础。
更新日期:2024-10-14
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