Department of Molecular Biosciences, Northwestern University, Evanston, United States.
NSF-Simons Center for Quantitative Biology, Northwestern University, Evanston, United States.
Elife. 2020 Jun 22;9:e56076. doi: 10.7554/eLife.56076.
Morphogen signaling contributes to the patterned spatiotemporal expression of genes during development. One mode of regulation of signaling-responsive genes is at the level of transcription. Single-cell quantitative studies of transcription have revealed that transcription occurs intermittently, in bursts. Although the effects of many gene regulatory mechanisms on transcriptional bursting have been studied, it remains unclear how morphogen gradients affect this dynamic property of downstream genes. Here we have adapted single molecule fluorescence in situ hybridization (smFISH) for use in the wing imaginal disc in order to measure nascent and mature mRNA of genes downstream of the Wg and Dpp morphogen gradients. We compared our experimental results with predictions from stochastic models of transcription, which indicated that the transcription levels of these genes appear to share a common method of control via burst frequency modulation. Our data help further elucidate the link between developmental gene regulatory mechanisms and transcriptional bursting.
形态发生素信号在发育过程中有助于基因的有图案的时空表达。信号响应基因的一种调控模式是在转录水平上。对转录的单细胞定量研究表明,转录是间歇性的,呈爆发式进行。尽管许多基因调控机制对转录爆发的影响已经被研究过,但仍不清楚形态发生素梯度如何影响下游基因的这种动态特性。在这里,我们采用单分子荧光原位杂交(smFISH)技术,用于研究 Wg 和 Dpp 形态发生素梯度下游基因的新生和成熟 mRNA。我们将实验结果与转录的随机模型的预测进行了比较,结果表明,这些基因的转录水平似乎通过爆发频率调制共享一种共同的控制方法。我们的数据有助于进一步阐明发育基因调控机制与转录爆发之间的联系。