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Calmodulin-mediated signal transduction pathways in Arabidopsis are fine-tuned by methylation.

机译:拟南芥中钙调蛋白介导的信号转导途径可通过甲基化进行微调。

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摘要

Calmodulin N-methyltransferase (CaM KMT) is an evolutionarily conserved enzyme in eukaryotes that transfers three methyl groups to a highly conserved lysyl residue at position 115 in calmodulin (CaM). We sought to elucidate whether the methylation status of CaM plays a role in CaM-mediated signaling pathways by gene expression analyses of CaM KMT and phenotypic characterization of Arabidopsis thaliana lines wherein CaM KMT was overexpressed (OX), partially silenced, or knocked out. CaM KMT was expressed in discreet spatial and tissue-specific patterns, most notably in root tips, floral buds, stamens, apical meristems, and germinating seeds. Analysis of transgenic plants with genetic dysfunction in CaM KMT revealed a link between the methylation status of CaM and root length. Plants with suppressed CaM methylation had longer roots and CaM KMT OX lines had shorter roots than wild type (Columbia-0). CaM KMT was also found to influence the root radial developmental program. Protein microarray analyses revealed a number of proteins with specificity for methylated forms of CaM, providing candidate functional intermediates between the observed phenotypes and the target pathways. This work demonstrates that the functionality of the large CaM family in plants is fine-tuned by an overarching methylation mechanism.
机译:钙调蛋白N-甲基转移酶(CaM KMT)是真核生物中一种进化保守的酶,可将三个甲基转移至钙调蛋白(CaM)115位的高度保守的赖氨酰残基。我们试图通过对CaM KMT的基因表达分析和拟南芥其中CaM KMT过表达(OX),部分沉默或敲除的拟南芥系的表型表征,来阐明CaM的甲基化状态是否在CaM介导的信号通路中起作用。 CaM KMT以离散的空间和组织特定模式表达,最明显的是在根尖,花蕾,雄蕊,顶端分生组织和发芽种子中表达。对CaM KMT中具有遗传功能障碍的转基因植物的分析表明,CaM的甲基化状态与根长之间存在联系。与野生型(Columbia-0)相比,具有受抑制的CaM甲基化作用的植物具有更长的根,而CaM KMT OX系具有短的根。还发现CaM KMT影响根部径向发育计划。蛋白质微阵列分析揭示了许多对CaM的甲基化形式具有特异性的蛋白质,在观察到的表型和靶标通路之间提供了候选功能中间体。这项工作表明,植物中大型CaM家族的功能通过总体甲基化机制进行了微调。

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