Goossens Jonas, Swinnen Gwen, Vanden Bossche Robin, Pauwels Laurens, Goossens Alain
Department of Plant Systems Biology, Flanders Institute for Biotechnology (VIB), B-9052, Gent, Belgium.
Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052, Gent, Belgium.
New Phytol. 2015 Jun;206(4):1229-37. doi: 10.1111/nph.13398. Epub 2015 Mar 27.
The bHLH transcription factor MYC2, together with its paralogues MYC3 and MYC4, is a master regulator of the response to the jasmonate (JA) hormone in Arabidopsis (Arabidopsis thaliana). In the absence of JA, JASMONATE ZIM (JAZ) proteins interact with the MYC proteins to block their activity. Understanding of the mechanism and specificity of this interaction is key to unravel JA signalling. We generated mutant MYC proteins and assessed their activity and the specificity of their interaction with the 12 Arabidopsis JAZ proteins. We show that the D94N mutation present in the atr2D allele of MYC3 abolishes the interaction between MYC3 and most JAZ proteins. The same effect is observed when the corresponding conserved Asp (D105) was mutated in MYC2. Accordingly, MYC2(D105N) activated target genes in the presence of JAZ proteins, in contrast to wild-type MYC2. JAZ1 and JAZ10 were the only JAZ proteins still showing interaction with the mutant MYC proteins, due to a second MYC interaction domain, besides the classical Jas domain. Our results visualize the divergence among JAZ proteins in their interaction with MYC proteins. Ultimately, the transferability of the Asp-to-Asn amino acid change might facilitate the design of hyperactive transcription factors for plant engineering.
bHLH转录因子MYC2与其旁系同源蛋白MYC3和MYC4一起,是拟南芥中茉莉酸(JA)激素应答的主要调节因子。在没有JA的情况下,茉莉酸ZIM(JAZ)蛋白与MYC蛋白相互作用以阻断其活性。了解这种相互作用的机制和特异性是揭示JA信号传导的关键。我们生成了突变型MYC蛋白,并评估了它们的活性以及与12种拟南芥JAZ蛋白相互作用的特异性。我们发现,MYC3的atr2D等位基因中存在的D94N突变消除了MYC3与大多数JAZ蛋白之间的相互作用。当MYC2中相应的保守天冬氨酸(D105)发生突变时,也观察到了相同的效果。因此,与野生型MYC2不同,MYC2(D105N)在存在JAZ蛋白的情况下激活了靶基因。JAZ1和JAZ10是仅有的仍与突变型MYC蛋白显示相互作用的JAZ蛋白,这是由于除了经典的Jas结构域之外还有第二个MYC相互作用结构域。我们的结果揭示了JAZ蛋白在与MYC蛋白相互作用方面的差异。最终,从天冬氨酸到天冬酰胺的氨基酸变化的可转移性可能有助于设计用于植物工程的高活性转录因子。