Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy
Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy
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DOI：
10.7554/elife.14770
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发表时间：
2016-06-11
期刊：
影响因子：
7.7
通讯作者：
Sozzani, Rosangela
中科院分区：
文献类型：
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作者：
Clark, Natalie M.;Hinde, Elizabeth;Sozzani, Rosangela
为了理解多细胞生物中复杂的调控过程，能够定量分析蛋白质运动和蛋白质-蛋白质相互作用在时间和空间上是至关重要的。在拟南芥发育过程中，短根（SHORTROOT， SHR）的细胞间运动及其与下游靶点稻草人（稻草人，SCR）的相互作用控制着根的形态和细胞命运的规范。然而，目前还没有关于SHR运动和SHR- scr相互作用的时空动态定量信息。在这里，我们使用荧光相关光谱（FCS）技术组合量化参数，包括SHR迁移率，寡聚态以及与SCR的关联。然后，我们将这些参数纳入到SHR和SCR的数学模型中，结果表明SHR在几分钟内达到稳态，而SCR和SHR-SCR复合物在18 - 24小时内达到稳态。我们的模型揭示了SHR和SCR动力学的时间，并使我们能够理解蛋白质运动和蛋白质化学计量学如何促进发育。
To understand complex regulatory processes in multicellular organisms, it is critical to be able to quantitatively analyze protein movement and protein-protein interactions in time and space. During Arabidopsis development, the intercellular movement of SHORTROOT (SHR) and subsequent interaction with its downstream target SCARECROW (SCR) control root patterning and cell fate specification. However, quantitative information about the spatio-temporal dynamics of SHR movement and SHR-SCR interaction is currently unavailable. Here, we quantify parameters including SHR mobility, oligomeric state, and association with SCR using a combination of Fluorescent Correlation Spectroscopy (FCS) techniques. We then incorporate these parameters into a mathematical model of SHR and SCR, which shows that SHR reaches a steady state in minutes, while SCR and the SHR-SCR complex reach a steady-state between 18 and 24 hr. Our model reveals the timing of SHR and SCR dynamics and allows us to understand how protein movement and protein-protein stoichiometry contribute to development.