Glutamate signalling via a MEKK1 kinase-dependent pathway induces changes in Arabidopsis root architecture.
Glutamate signalling via a MEKK1 kinase-dependent pathway induces changes in Arabidopsis root architecture.
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DOI：
10.1111/tpj.12201
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发表时间：
2013-07
期刊：
影响因子：
--
通讯作者：
Krysan PJ
中科院分区：
文献类型：
--
作者：
Forde BG;Cutler SR;Zaman N;Krysan PJ
利用化学遗传学方法研究了外源谷氨酸（l-Glu）对拟南芥根构型的影响。使用专门开发的96孔微表型分析系统，对哺乳动物谷氨酸和GABA受体的80种激动剂和拮抗剂进行了初步筛选，发现没有一种能复制根部对l-Glu的反应或拮抗l-Glu。然而，使用酿酒酵母（酵母）中超过1500种生物活性分子进行的更大筛选确定了两组干扰l-Glu反应的分子。其中一种拮抗剂，2-（4-氯-3-甲基苯基）-2-氧代乙基硫氰酸酯（CMOT），已被报道靶向Ste 11，一种在酵母中进化保守的MAP激酶（MAP 3 K）。这导致发现，根生长在三重mekk 1 mekk 2 mekk 3突变体（mekk 1/2/3），缺陷的一组三个串联排列的MAP 3 Ks，几乎是不敏感的l-Glu。然而，mekk 1/2/3根的敏感性抑制其他氨基酸的谷氨酸受体样（GLR）通道的激动剂在拟南芥根（天冬酰胺，半胱氨酸，甘氨酸和丝氨酸）的报道不受影响。通过用携带完整MEKK 1基因的构建体转化，恢复了mekk 1/2/3突变体的L-Glu敏感性。这些结果表明，MEKK 1在转导L-Glu信号中起着关键作用，该信号引起根结构的大规模变化，并为植物中存在类似于动物中发现的L-Glu信号通路提供遗传证据。
A chemical genetic approach has been used to investigate the mechanism by which external glutamate (l–Glu) is able to trigger major changes in root architecture in Arabidopsis thaliana L. An initial screen of 80 agonists and antagonists of mammalian glutamate and GABA receptors, using a specially developed 96-well microphenotyping system, found none that replicated the response of the root to l–Glu or antagonized it. However, a larger screen using >1500 molecules bioactive in Saccharomyces cerevisiae (yeast) identified two groups that interfered with the l–Glu response. One of the antagonists, 2–(4–chloro-3-methylphenyl)-2-oxoethyl thiocyanate (CMOT), has been reported to target Ste11, an evolutionarily conserved MAP kinase kinase kinase (MAP3K) in yeast. This led to the discovery that root growth in a triple mekk1 mekk2 mekk3 mutant (mekk1/2/3), defective in a set of three tandemly arranged MAP3Ks, was almost insensitive to l–Glu. However, the sensitivity of mekk1/2/3 roots to inhibition by other amino acids reported to act as agonists of glutamate receptor-like (GLR) channels in Arabidopsis roots (Asn, Cys, Gly and Ser) was unaffected. The l–Glu sensitivity of the mekk1/2/3 mutant was restored by transformation with a construct carrying the intact MEKK1 gene. These results demonstrate that MEKK1 plays a key role in transducing the l–Glu signal that elicits large-scale changes in root architecture, and provide genetic evidence for the existence in plants of an l–Glu signalling pathway analogous to that found in animals.
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