Wilson K, Long D, Swinburne J, Coupland G
Department of Molecular Genetics, John Innes Centre, Colney, Norwich, United Kingdom.
Plant Cell. 1996 Apr;8(4):659-71. doi: 10.1105/tpc.8.4.659.
A novel transposon-tagging strategy designed to recover dominant gain-of-function alleles was performed with Arabidopsis by using a Dissociation element with a cauliflower mosaic virus 35S promoter transcribing outward over one terminus. Lines containing transposed copies of this transposon were screened for mutants, and a semidominant mutation affecting plant height, hypocotyl elongation, and fertility was recovered. The pleiotropic effects of this mutation appear to result from a general reduction in cell expansion, and some of the effects are similar to those caused by supplying exogenous ethylene or cytokinin to wild-type seedlings. In addition, the arrangement of cells in some organs such as the etiolated hypocotyl, is disorganized. The mutation was called tiny, and the affected gene was cloned by first using transposon sequences to isolate the mutant allele. The predicted protein product of the TINY gene shows strong homology with the DNA binding domain of a recently identified class of plant transcription factors. This domain, called the APETALA2 domain, was initially identified as a duplicated region within the APETALA2 gene of Arabidopsis and then as a conserved region between APETALA2 and the ethylene responsive element binding proteins of tobacco. In the mutant allele, the Dissociation element is inserted in the untranslated leader of the TINY gene, 36 bp from the ATG, and the mutant contains a novel transcript that initiates from the cauliflower mosaic virus 35S promoter within the transposon. This transcript is present in greater abundance than the wild-type TINY transcript; therefore, the semidominant tiny mutation most likely results from increased, or ectopic, expression of the gene.
利用一个带有花椰菜花叶病毒35S启动子且在一个末端向外转录的解离元件,在拟南芥中实施了一种旨在获得显性功能获得性等位基因的新型转座子标签策略。筛选含有该转座子转座拷贝的株系以寻找突变体,获得了一个影响株高、下胚轴伸长和育性的半显性突变。该突变的多效性效应似乎是由细胞扩张普遍减少导致的,其中一些效应类似于向野生型幼苗供应外源乙烯或细胞分裂素所引起的效应。此外,在一些器官如黄化下胚轴中,细胞排列紊乱。该突变被命名为tiny,通过首先利用转座子序列分离突变等位基因来克隆受影响的基因。TINY基因的预测蛋白质产物与最近鉴定出的一类植物转录因子的DNA结合结构域具有很强的同源性。这个结构域称为APETALA2结构域,最初被鉴定为拟南芥APETALA2基因内的一个重复区域,随后被鉴定为APETALA2与烟草乙烯反应元件结合蛋白之间的一个保守区域。在突变等位基因中,解离元件插入到TINY基因的非翻译前导序列中,距离ATG为36 bp,并且该突变体包含一个从转座子内的花椰菜花叶病毒35S启动子起始的新转录本。这个转录本的丰度高于野生型TINY转录本;因此,半显性的tiny突变很可能是由该基因表达增加或异位表达导致的。