de Vetten N, Quattrocchio F, Mol J, Koes R
Department of Genetics, Institute for Molecular Biological Sciences, Vrije Universiteit, BioCentrum Amsterdam, The Netherlands.
Genes Dev. 1997 Jun 1;11(11):1422-34. doi: 10.1101/gad.11.11.1422.
In petunia flowers, the loci an1, an2, and an11 control the pigmentation of the flower by stimulating the transcription of anthocyanin biosynthetic genes. The an1 and an2 locus were recently cloned and encode a basic helix-loop-helix (bHLH) and MYB-domain transcriptional activator, respectively. Here, we report the isolation of the an11 locus by transposon tagging. RNA gel blot experiments show that an11 is expressed independently from an1 and an2 throughout plant development, as well as in tissues that do not express the anthocyanin pathway. It encodes a novel WD-repeat protein that is highly conserved even in species that do not produce anthocyanins such as yeast, nematodes, and mammals. The observation that the human an11 homolog partially complements the an11 petunia mutant in transient assays shows that sequence similarity reflects functional conservation. Overexpression of an2 in an11- petals restored the activity of a structural anthocyanin gene in transient assays, indicating that AN11 acts upstream of AN2. Cell fractionation experiments show that the bulk of the AN11 protein is localized in the cytoplasm. Taken together, this indicates that AN11 is a cytoplasmic component of a conserved signal transduction cascade that modulates AN2 function in petunia, thereby linking cellular signals with transcriptional activation.
在矮牵牛属花卉中,an1、an2和an11基因座通过刺激花青素生物合成基因的转录来控制花朵的色素沉着。an1和an2基因座最近已被克隆,分别编码一种碱性螺旋-环-螺旋(bHLH)和MYB结构域转录激活因子。在此,我们报道了通过转座子标签法分离出an11基因座。RNA凝胶印迹实验表明,an11在整个植物发育过程中以及在不表达花青素途径的组织中独立于an1和an2表达。它编码一种新型WD重复蛋白,这种蛋白即使在不产生花青素的物种(如酵母、线虫和哺乳动物)中也高度保守。在瞬时分析中,人类an11同源物部分互补矮牵牛an11突变体的观察结果表明,序列相似性反映了功能保守性。在瞬时分析中,在an11花瓣中过表达an2可恢复结构花青素基因的活性,这表明AN11在AN2的上游起作用。细胞分级分离实验表明,大部分AN11蛋白定位于细胞质中。综上所述,这表明AN11是一个保守信号转导级联反应的细胞质成分,该级联反应调节矮牵牛中AN2的功能,从而将细胞信号与转录激活联系起来。