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Characterization of the Class IV Homeodomain-Leucine Zipper Gene Family in Arabidopsis1,[W]

机译:拟南芥中IV类同源域-亮氨酸拉链基因家族的特征[W]

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The Arabidopsis (Arabidopsis thaliana) genome contains 16 genes belonging to the class IV homeodomain-Leucine zipper gene family. These include GLABRA2, ANTHOCYANINLESS2, FWA, ARABIDOPSIS THALIANA MERISTEM LAYER1 (ATML1), and PROTODERMAL FACTOR2 (PDF2). Our previous study revealed that atml1 pdf2 double mutants have severe defects in the shoot epidermal cell differentiation. Here, we have characterized additional members of this gene family, which we designated HOMEODOMAIN GLABROUS1 (HDG1) through HDG12. Analyses of transgenic Arabidopsis plants carrying the gene-specific promoter fused to the bacterial -glucuronidase reporter gene revealed that some of the promoters have high activities in the epidermal layer of the shoot apical meristem and developing shoot organs, while others are temporarily active during reproductive organ development. Expression profiles of highly conserved paralogous gene pairs within the family were found to be not necessarily overlapping. Analyses of T-DNA insertion mutants of these HDG genes revealed that all mutants except hdg11 alleles exhibit no abnormal phenotypes. hdg11 mutants show excess branching of the trichome. This phenotype is enhanced in hdg11 hdg12 double mutants. Double mutants were constructed for other paralogous gene pairs and genes within the same subfamily. However, novel phenotypes were observed only for hdg3 atml1 and hdg3 pdf2 mutants that both exhibited defects in cotyledon development. These observations suggest that some of the class IV homeodomain-Leucine zipper members act redundantly with other members of the family during various aspects of cell differentiation. DNA-binding sites were determined for two of the family members using polymerase chain reaction-assisted DNA selection from random oligonucleotides with their recombinant proteins. The binding sites were found to be similar to those previously identified for ATML1 and PDF2, which correspond to the pseudopalindromic sequence 5'-GCATTAAATGC-3' as the preferential binding site.
机译:拟南芥(Arabidopsis thaliana)基因组包含16个属于IV类同源域-亮氨酸拉链基因家族的基因。其中包括GLABRA2,ANTHOCYANINLESS2,FWA,阿拉伯拟南芥分离层1(ATML1)和原皮因子2(PDF2)。我们先前的研究表明atml1 pdf2双重突变体在茎表皮细胞分化中具有严重缺陷。在这里,我们已经表征了该基因家族的其他成员,我们将其命名为HOMEODOMAIN GLABROUS1(HDG1)至HDG12。对携带融合有细菌-葡糖醛酸糖苷酶报道基因的基因特异性启动子的转基因拟南芥植物的分析表明,一些启动子在茎尖分生组织的表皮层和发育中的芽器官中具有高活性,而其他启动子在生殖器官中暂时活跃发展。发现该家族中高度保守的同源基因对的表达谱不一定重叠。这些HDG基因的T-DNA插入突变体的分析表明,除hdg11等位基因外,所有突变体均未显示异常表型。 hdg11突变体显示了毛状体的过度分支。此表型在hdg11 hdg12双重突变体中得到增强。为其他同源基因对和同一亚科中的基因构建了双突变体。但是,仅对于均在子叶发育中均表现出缺陷的hdg3 atml1和hdg3 pdf2突变体观察到了新的表型。这些观察结果表明,在细胞分化的各个方面,某些IV类同源域-亮氨酸拉链成员与该家族的其他成员有多余的作用。使用聚合酶链反应辅助DNA选择随机寡核苷酸及其重组蛋白,确定两个家族成员的DNA结合位点。发现结合位点类似于先前针对ATML1和PDF2鉴定的结合位点,其对应于假回文序列5'-GCATTAAATGC-3'作为优先结合位点。

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