开花植物 LIM 蛋白基因家族的分子和表达分析

A Eliasson, N Gass, C Mundel, R Baltz, R Kräuter, J L Evrard, A Steinmetz
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引用次数: 57

摘要

在真核生物中,LIM-结构域蛋白参与重要的细胞过程,包括基因转录和肌动蛋白细胞骨架组织。它们主要存在于动物中,但在酵母和植物中也有发现。在鉴定了向日葵花粉中的一种 LIM-结构域蛋白之后,我们对开花植物中的此类蛋白进行了广泛的研究。我们从向日葵、烟草和拟南芥中分离并研究了两种新型 LIM-结构域蛋白的 cDNA 和/或基因组序列。这些植物蛋白在结构上与动物中与细胞骨架相关的 CRP 类 LIM 蛋白有关,但显示出一些独特的特征,包括第二个非典型的 LIM 结构域。我们对这些基因、另外一个烟草基因和两个拟南芥基因(其序列可从数据库中获得)进行了比较表达研究。这些研究是在基因特异性引物作用下通过 RT-PCR 进行的,结果表明向日葵和烟草的花粉粒和孢子体组织表达不同的 LIM 蛋白。除了拟南芥的一个基因有两个内含子外,所有被分析的基因在保守的位置上都有四个内含子,这表明在高等植物中进化出不同拷贝的祖先基因已经具有这种分裂结构。
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Molecular and expression analysis of a LIM protein gene family from flowering plants.

LIM-domain proteins participate in important cellular processes in eukaryotes, including gene transcription and actin cytoskeleton organization. They are predominantly found in animals, but have also been identified in yeast and plants. Following the characterization ofa LIM-domain protein in sunflower pollen, we carried out an extensive search for these proteins in flowering plants. We have isolated and studied cDNAs and/or genomic sequences for two novel LIM-domain proteins from sunflower, three from tobacco, and one from Arabidopsis. The plant proteins are structurally related to the cytoskeleton-associated CRP class of LIM proteins in animals, but show several distinctive features, including a second, atypical, LIM domain. We have performed comparative expression studies of these genes, as well as of one other gene from tobacco and two additional Arabidopsis genes whose sequences are available from databases. These studies, carried out by RT-PCR in the presence of gene-specific primers, showed that, in sunflower and tobacco, pollen grains and sporophytic tissues express different sets of LIM proteins. With the exception of one Arabidopsis gene--which has two introns--all the genes analyzed contain four introns at conserved positions, indicating that the ancestral gene from which the various copies evolved in higher plants allready had this split structure.

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