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Creation of Master Transcription Factors to Regulate Plant Metabolic Pathways

机译:创建主要转录因子来调节植物代谢途径

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摘要

Key steps of biosynthetic pathways of plant secondary metabolites are regulated by multiple transcription factors (TFs). The ability to create single "master" TFs capable of regulating the expression of all genes in a particular pathway would be a powerful tool for metabolic pathway engineering. To test the feasibility, directed evolution technologies were applied to modify the properties of a family of plant bHLH-like TFs involved in flavonoid biosynthesis. Directed evolution utilizes genomic information and incorporates Darwinian principles of mutation and selection into experimental strategies for improving gene and protein functions. We have demonstrated that the transcriptional activity and promoter specificity of the bHLH TFs can be improved through recursive evolution. TF variants with increased transactivational activity and broadened promoter specificity have been generated. Ectopic expression of an evolved bHLH TF in transgenic tobacco resulted in increased anthocyanin accumulation compared to plants expressing the wild-type gene. Analysis of the genetic diversity of the evolved variants reveals the potential cooperation between functional domains of the bHLH TFs. Our study substantiates the potential of utilizing master TFs for regulation of important traits of plants.
机译:植物次生代谢产物生物合成途径的关键步骤受多种转录因子(TFs)调控。创建能够调节特定途径中所有基因表达的单个“主” TF的能力将是代谢途径工程的有力工具。为了测试可行性,应用定向进化技术来修饰参与类黄酮生物合成的植物bHLH样TF家族的特性。定向进化利用基因组信息并将突变和选择的达尔文原理结合到改善基因和蛋白质功能的实验策略中。我们已经证明,bHLH TFs的转录活性和启动子特异性可以通过递归进化得到改善。已经产生了具有增加的反式激活活性和拓宽的启动子特异性的TF变体。与表达野生型基因的植物相比,在转基因烟草中异位表达的bHLH TF的表达导致花青素积累增加。进化变异的遗传多样性分析揭示了bHLH TFs功能域之间的潜在合作。我们的研究证实了利用主TF调控植物重要性状的潜力。

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