Quantitative imaging of RNA polymerase II activity in plants reveals the single-cell basis of tissue-wide transcriptional dynamics.
Quantitative imaging of RNA polymerase II activity in plants reveals the single-cell basis of tissue-wide transcriptional dynamics.
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DOI：
10.1038/s41477-021-00976-0
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发表时间：
2021-08
期刊：
影响因子：
18
通讯作者：
Garcia HG
中科院分区：
文献类型：
--
作者：
Alamos S;Reimer A;Niyogi KK;Garcia HG
植物对环境的反应往往取决于转录调控的时空动态。虽然活体成像工具已被广泛用于定量捕获活动物细胞中的快速转录动态，但在植物中缺乏这些技术的实施限制了伴随的定量研究。在这里，我们应用PP 7和MS 2 RNA标记技术对活体植物单细胞对实验处理做出反应时的RNA聚合酶II活性动态进行定量成像。使用这种技术，我们计数新生RNA转录本实时烟草（烟草）和拟南芥（拟南芥）。热休克报告员的检查表明，植物组织通过调节参与转录的细胞数量而不是活性细胞的转录速率来响应外部信号。这种类似开关的行为，加上转录速率的细胞与细胞之间的变异性，导致mRNA产生的变异性跨越三个数量级。我们确定细胞异质性主要源于个体等位基因内在的随机性。两者合计，我们的研究结果表明，现在可以定量研究活植物单细胞中转录程序的动态。
The responses of plants to their environment often hinge on the spatiotemporal dynamics of transcriptional regulation. While live-imaging tools have been used extensively to quantitatively capture rapid transcriptional dynamics in living animal cells, lack of implementation of these technologies in plants has limited concomitant quantitative studies. Here, we applied the PP7 and MS2 RNA-labeling technologies for the quantitative imaging of RNA polymerase II activity dynamics in single cells of living plants as they respond to experimental treatments. Using this technology, we count nascent RNA transcripts in real-time in Nicotiana benthamiana (tobacco) and Arabidopsis thaliana (Arabidopsis). Examination of heat shock reporters revealed that plant tissues respond to external signals by modulating the number of cells engaged in transcription rather than the transcription rate of active cells. This switch-like behavior, combined with cell-to-cell variability in transcription rate, results in mRNA production variability spanning three orders of magnitude. We determined that cellular heterogeneity stems mainly from the stochasticity intrinsic to individual alleles. Taken together, our results demonstrate that it is now possible to quantitatively study the dynamics of transcriptional programs in single cells of living plants.
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影响因子：
11.1
作者：
Birnbaum KD
通讯作者：
Birnbaum KD
DOI：
10.1105/tpc.20.00439
发表时间：
2020-10
期刊：
The Plant cell
影响因子：
--
作者：
Iwatate RJ;Yoshinari A;Yagi N;Grzybowski M;Ogasawara H;Kamiya M;Komatsu T;Taki M;Yamaguchi S;Frommer WB;Nakamura M
通讯作者：
Nakamura M
影响因子：
5.1
作者：
Duncan S;Olsson TSG;Hartley M;Dean C;Rosa S
通讯作者：
Rosa S
影响因子：
16.6
作者：
Araújo IS;Pietsch JM;Keizer EM;Greese B;Balkunde R;Fleck C;Hülskamp M
通讯作者：
Hülskamp M
影响因子：
9.3
作者：
Ietswaart R;Rosa S;Wu Z;Dean C;Howard M
通讯作者：
Howard M