Dynamic Cell Imaging, University of Bielefeld, Bielefeld, Germany.
PLoS One. 2013;8(1):e53737. doi: 10.1371/journal.pone.0053737. Epub 2013 Jan 11.
In the present work, the objective has been to analyse the compatibility of plant and human transcriptional machinery. The experiments revealed that nuclear import and export are conserved among plants and mammals. Further it has been shown that transactivation of a human promoter occurs by human transcription factor NF-κB in plant cells, demonstrating that the transcriptional machinery is highly conserved in both kingdoms. Functionality was also seen for regulatory elements of NF-κB such as its inhibitor IκB isoform α that negatively regulated the transactivation activity of the p50/RelA heterodimer by interaction with NF-κB in plant cells. Nuclear export of RelA could be demonstrated by FRAP-measurements so that RelA shows nucleo-cytoplasmic shuttling as reported for RelA in mammalian cells. The data reveals the high level of compatibility of human transcriptional elements with the plant transcriptional machinery. Thus, Arabidopsis thaliana mesophyll protoplasts might provide a new heterologous expression system for the investigation of the human NF-κB signaling pathways. The system successfully enabled the controlled manipulation of NF-κB activity. We suggest the plant protoplast system as a tool for reconstitution and analyses of mammalian pathways and for direct observation of responses to e.g. pharmaceuticals. The major advantage of the system is the absence of interference with endogenous factors that affect and crosstalk with the pathway.
在本工作中,我们的目标是分析植物和人类转录机制的兼容性。实验表明,核输入和输出在植物和哺乳动物中是保守的。进一步表明,人类转录因子 NF-κB 在植物细胞中激活了人类启动子,证明了这两个领域的转录机制具有高度的保守性。NF-κB 的调节元件,如其抑制剂 IκB 同工型 α,也具有功能,通过与植物细胞中的 NF-κB 相互作用,负调控 p50/RelA 异二聚体的转录激活活性。通过 FRAP 测量可以证明 RelA 的核输出,因此 RelA 显示出核质穿梭,正如在哺乳动物细胞中报道的 RelA 一样。这些数据揭示了人类转录元件与植物转录机制的高度兼容性。因此,拟南芥叶肉原生质体可能为研究人类 NF-κB 信号通路提供了一种新的异源表达系统。该系统成功地实现了对 NF-κB 活性的控制操作。我们建议将植物原生质体系统作为用于重建和分析哺乳动物途径以及直接观察对例如药物的反应的工具。该系统的主要优点是不存在干扰途径的内源性因素。