Kandasamy Muthugapatti K, Deal Roger B, McKinney Elizabeth C, Meagher Richard B
Department of Genetics, Life Sciences Building, University of Georgia, Athens, GA 30602, USA.
Plant J. 2005 Mar;41(6):845-58. doi: 10.1111/j.1365-313X.2005.02345.x.
Actin-related proteins (ARPs) share moderate sequence homology and basal structure with conventional actins and are found in all eukaryotes. While the functions of most of the divergent ARPs are not clear, several of them are localized to the nucleus and have been identified as components of various chromatin-modifying complexes. Using an antibody to Arabidopsis AtARP4, we found this conserved homolog of human BAF53 and yeast Arp4 is concentrated in the nucleoplasm of Arabidopsis, Brassica, and tobacco cells. To gain further insight into the role of ARP4, we have examined Arabidopsis plants that are defective in AtARP4 expression. Phenotypic analysis of the arp4-1 mutant allele, which has a T-DNA insertion in the promoter region and a moderate reduction in the level of AtARP4 protein expression, revealed partial sterility due to defects in anther development. Targeting the distinct, 3' UTR of AtARP4 transcripts with RNA interference caused a drastic reduction in the level of AtARP4 protein expression in several independent transgenic lines, and resulted in strong pleiotropic phenotypes such as altered organization of plant organs, early flowering, delayed flower senescence and high levels of sterility. Western blot analysis and immunolabeling demonstrated a clear correlation between reductions in the level of AtARP4 expression and severity of the phenotypes. Based on our results and data on the orthologs of AtARP4 in yeast and other organisms, we suggest that AtARP4 is likely to exert its effects on plant development through the modulation of chromatin structure and subsequent changes in gene regulation.
肌动蛋白相关蛋白(ARPs)与传统肌动蛋白具有适度的序列同源性和基本结构,存在于所有真核生物中。虽然大多数不同的ARPs的功能尚不清楚,但其中一些定位于细胞核,并已被鉴定为各种染色质修饰复合物的组成部分。使用针对拟南芥AtARP4的抗体,我们发现人类BAF53和酵母Arp4的这种保守同源物集中在拟南芥、甘蓝和烟草细胞的核质中。为了进一步深入了解ARP4的作用,我们研究了AtARP4表达有缺陷的拟南芥植株。对arp4 - 1突变等位基因进行表型分析,该等位基因在启动子区域有一个T-DNA插入,AtARP4蛋白表达水平适度降低,结果显示由于花药发育缺陷导致部分不育。用RNA干扰靶向AtARP4转录本独特的3' UTR,导致几个独立转基因系中AtARP4蛋白表达水平急剧降低,并导致强烈的多效性表型,如植物器官组织改变、早花、花衰老延迟和高度不育。蛋白质免疫印迹分析和免疫标记表明AtARP4表达水平的降低与表型严重程度之间存在明显的相关性。基于我们的结果以及酵母和其他生物体中AtARP