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A chemical genetic screen reveals that iminosugar inhibitors of plant glucosylceramide synthase inhibit root growth in Arabidopsis and cereals.
Scientific Reports ( IF 3.8 ) Pub Date : 2018-Nov-06 , DOI: 10.1038/s41598-018-34749-1
Michael D Rugen 1 , Mathieu M J L Vernet 1 , Laila Hantouti 1 , Amalia Soenens 2, 3 , Vasilios M E Andriotis 2, 4 , Martin Rejzek 1 , Paul Brett 2 , Richard J B H N van den Berg 5 , Johannes M F G Aerts 6 , Hermen S Overkleeft 5 , Robert A Field 1
Affiliation  

Iminosugars are carbohydrate mimics that are useful as molecular probes to dissect metabolism in plants. To analyse the effects of iminosugar derivatives on germination and seedling growth, we screened a library of 390 N-substituted iminosugar analogues against Arabidopsis and the small cereal Eragrostis tef (Tef). The most potent compound identified in both systems, N-5-(adamantane-1-yl-ethoxy)pentyl- L-ido-deoxynojirimycin (L-ido-AEP-DNJ), inhibited root growth in agar plate assays by 92% and 96% in Arabidopsis and Tef respectively, at 10 µM concentration. Phenocopying the effect of L-ido-AEP-DNJ with the commercial inhibitor (PDMP) implicated glucosylceramide synthase as the target responsible for root growth inhibition. L-ido-AEP-DNJ was twenty-fold more potent than PDMP. Liquid chromatography-mass spectrometry (LC-MS) analysis of ceramide:glucosylceramide ratios in inhibitor-treated Arabidopsis seedlings showed a decrease in the relative quantity of the latter, confirming that glucosylceramide synthesis is perturbed in inhibitor-treated plants. Bioinformatic analysis of glucosylceramide synthase indicates gene conservation across higher plants. Previous T-DNA insertional inactivation of glucosylceramide synthase in Arabidopsis caused seedling lethality, indicating a role in growth and development. The compounds identified herein represent chemical alternatives that can overcome issues caused by genetic intervention. These inhibitors offer the potential to dissect the roles of glucosylceramides in polyploid crop species.

中文翻译:


化学遗传筛选表明,植物葡萄糖神经酰胺合酶的亚氨基糖抑制剂可抑制拟南芥和谷物的根部生长。



亚氨基糖是碳水化合物模拟物,可用作剖析植物代谢的分子探针。为了分析亚氨基糖衍生物对发芽和幼苗生长的影响,我们筛选了针对拟南芥和小谷类画眉草 tef (Tef) 的 390 个 N 取代亚氨基糖类似物文库。在这两个系统中鉴定出的最有效的化合物 N-5-(金刚烷-1-基-乙氧基)戊基-L-ido-脱氧野尻霉素 (L-ido-AEP-DNJ) 在琼脂平板试验中抑制根生长达 92%,拟南芥和 Tef 中浓度为 10 µM 时分别为 96%。用商业抑制剂 (PDMP) 对 L-ido-AEP-DNJ 的效果进行表型复制,表明葡萄糖神经酰胺合酶是抑制根生长的靶标。 L-ido-AEP-DNJ 的效力比 PDMP 强二十倍。对抑制剂处理的拟南芥幼苗中的神经酰胺:葡萄糖神经酰胺比率进行液相色谱-质谱(LC-MS)分析,结果显示后者的相对数量减少,证实了抑制剂处理的植物中葡萄糖神经酰胺的合成受到干扰。葡萄糖神经酰胺合酶的生物信息分析表明高等植物中的基因保守性。此前,拟南芥中葡萄糖神经酰胺合酶的 T-DNA 插入失活导致幼苗死亡,表明其在生长和发育中发挥作用。本文鉴定的化合物代表了可以克服遗传干预引起的问题的化学替代品。这些抑制剂提供了剖析葡萄糖神经酰胺在多倍体作物物种中的作用的潜力。
更新日期:2018-11-06
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