Yang Yingzhen, Xiang Hongjun, Jack Thomas
Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.
Plant J. 2003 Jan;33(1):177-88. doi: 10.1046/j.1365-313x.2003.01603.x.
The Arabidopsis floral organ identity genes APETALA3 (AP3) and PISTILLATA (PI) encode related DNA-binding proteins of the MADS family. Considerable evidence supports the hypothesis that a heterodimer of AP3 and PI is an essential component of B class activity. All ap3 and pi alleles characterized to date exhibit equivalent phenotypic defects in both whorls 2 and 3. In strong ap3 and pi mutants, petals and stamens are missing and sepals and carpels develop in their place. Weak ap3 and pi mutants exhibit partial conversions of petals to sepals and stamens to carpels. In this report, we describe the isolation and characterization of pi-5, an unusual B class mutant that exhibits defects in whorl 2 where sepals develop in place of petals, but third whorl stamens are most often normal. pi-5 flowers resemble those from 35S::SEP3 antisense plants. pi-5 contains missense mutation in the K domain (PIE125K). PIE125K exhibits defects in heterodimerization with its partner protein AP3. Via a reverse yeast two-hybrid screen, AP3K139E was isolated as a compensatory mutant of PIE125K. The compensatory interaction between PIE125K and AP3K139E is observed both in yeast two-hybrid assays and in planta. On its own, AP3K139E exhibits defects in specifying both petal and stamen identity. In addition, PIE125K is defective in interaction with SEPALLATA proteins in both two- and three-hybrid assays suggesting that PIE125K is defective in forming higher order complexes of MADS proteins. The decreased concentration of PI/AP3/SEP complexes offers an explanation for the petal defects observed in both pi-5 and 35S::SEP3 antisense plants.
拟南芥花器官特征基因APETALA3(AP3)和PISTILLATA(PI)编码MADS家族中相关的DNA结合蛋白。大量证据支持以下假说:AP3和PI的异二聚体是B类活性的重要组成部分。迄今为止所鉴定的所有ap3和pi等位基因在第2轮和第3轮中均表现出相同的表型缺陷。在强ap3和pi突变体中,花瓣和雄蕊缺失,萼片和心皮取而代之发育。弱ap3和pi突变体表现出花瓣部分转变为萼片,雄蕊部分转变为心皮。在本报告中,我们描述了pi - 5的分离和特征,pi - 5是一种不寻常的B类突变体,其在第2轮中表现出缺陷,萼片取代花瓣发育,但第3轮雄蕊大多正常。pi - 5花类似于35S::SEP3反义植物的花。pi - 5在K结构域(PIE125K)中含有错义突变。PIE125K在与其伴侣蛋白AP3异二聚化时表现出缺陷。通过反向酵母双杂交筛选,分离出AP3K139E作为PIE125K的补偿突变体。在酵母双杂交试验和植物体内均观察到PIE125K与AP3K139E之间的补偿性相互作用。单独来看,AP3K139E在确定花瓣和雄蕊特征方面均表现出缺陷。此外,在双杂交和三杂交试验中,PIE125K与SEPALLATA蛋白相互作用存在缺陷,这表明PIE125K在形成MADS蛋白的高阶复合物方面存在缺陷。PI/AP3/SEP复合物浓度的降低为在pi - 5和35S::SEP3反义植物中观察到的花瓣缺陷提供了解释。