Multifunctional chemical inhibitors of the florigen activation?complex discovered by structure‐based high‐throughput screening
Multifunctional chemical inhibitors of the florigen activation?complex discovered by structure‐based high‐throughput screening
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通过基于结构的高通量筛选发现成花素激活复合物的多功能化学抑制剂
DOI：
10.1111/tpj.16008
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Kojima Chojiro
中科院分区：
文献类型：
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作者：
Taoka Ken‐ichiro;Kawahara Ikumi;Shinya Shoko;Harada Ken‐ichi;Yamashita Eiki;Shimatani Zenpei;Furuita Kyoko;Muranaka Tomoaki;Oyama Tokitaka;Terada Rie;Nakagawa Atsushi;Fujiwara Toshimichi;Tsuji Hiroyuki;Kojima Chojiro
基于结构的高通量筛选靶向蛋白质-蛋白质相互作用（PPI）的化合物是一种很有前途的技术，可以深入了解植物发育是如何调节的，从而导致许多潜在的农业应用。目前，还没有使用高通量筛选来鉴定靶向植物转录复合物的化学物质的例子，其中一些化合物负责调节多种生理功能。成花素是一种由FLOWERLOCUS T（FT）编码的蛋白质，最初被鉴定为促进开花的分子，此后被证明调节马铃薯（Solanum tuberosum）的开花和其他发育现象，如块茎的形成。FT与14 - 3 - 3支架蛋白和FD（一种激活下游基因表达的bZIP转录因子）一起作为成花素活化复合物（FAC）的组分发挥作用。虽然14 - 3 - 3是FAC的重要组成部分，但对14 - 3 - 3蛋白本身的功能知之甚少。在这里，我们报告了高通量的体外荧光共振能量转移（FRET）筛选化学文库的结果，使我们能够识别能够抑制FAC形成的小分子。这些分子通过与14 - 3 - 3蛋白直接结合，消除了14 - 3 - 3蛋白与OsFD 1肽（一种水稻FD）之间的体外相互作用。用特异性命中分子S4处理，以剂量依赖性方式强烈抑制FAC活性和浮萍中的开花、马铃薯中的块茎形成和水稻中的分枝。我们的研究结果表明，基于PPI三维结构的高通量筛选方法适用于植物。在这项研究中，我们提出了良好的候选化合物，用于未来的修饰，以通过抑制FAC形成来获得成花素依赖性过程的抑制剂。
Structure‐based high‐throughput screening of chemical compounds that target protein–protein interactions (PPIs) is a promising technology for gaining insight into how plant development is regulated, leading to many potential agricultural applications. At present, there are no examples of using high‐throughput screening to identify chemicals that target plant transcriptional complexes, some of which are responsible for regulating multiple physiological functions. Florigen, a protein encoded byFLOWERING LOCUS T(FT), was initially identified as a molecule that promotes flowering and has since been shown to regulate flowering and other developmental phenomena such as tuber formation in potato (Solanum tuberosum). FT functions as a component of the florigen activation complex (FAC) with a 14‐3‐3 scaffold protein and FD, a bZIP transcription factor that activates downstream gene expression. Although 14‐3‐3 is an important component of FAC, little is known about the function of the 14‐3‐3 protein itself. Here, we report the results of a high‐throughputin vitrofluorescence resonance energy transfer (FRET) screening of chemical libraries that enabled us to identify small molecules capable of inhibiting FAC formation. These molecules abrogate thein vitrointeraction between the 14‐3‐3 protein and the OsFD1 peptide, a rice (Oryza sativa) FD, by directly binding to the 14‐3‐3 protein. Treatment with S4, a specific hit molecule, strongly inhibited FAC activity and flowering in duckweed, tuber formation in potato, and branching in rice in a dose‐dependent manner. Our results demonstrate that the high‐throughput screening approach based on the three‐dimensional structure of PPIs is suitable in plants. In this study, we have proposed good candidate compounds for future modification to obtain inhibitors of florigen‐dependent processes through inhibition of FAC formation.