Yang Guojun, Weil Clifford F, Wessler Susan R
Department of Plant Biology, University of Georgia, Athens, Georgia 30602, USA.
Plant Cell. 2006 Oct;18(10):2469-78. doi: 10.1105/tpc.106.045906. Epub 2006 Oct 13.
The Tc1/mariner transposable element superfamily is widely distributed in animal and plant genomes. However, no active plant element has been previously identified. Nearly identical copies of a rice (Oryza sativa) Tc1/mariner element called Osmar5 in the genome suggested potential activity. Previous studies revealed that Osmar5 encoded a protein that bound specifically to its own ends. In this report, we show that Osmar5 is an active transposable element by demonstrating that expression of its coding sequence in yeast promotes the excision of a nonautonomous Osmar5 element located in a reporter construct. Element excision produces transposon footprints, whereas element reinsertion occurs at TA dinucleotides that were either tightly linked or unlinked to the excision site. Several site-directed mutations in the transposase abolished activity, whereas mutations in the transposase binding site prevented transposition of the nonautonomous element from the reporter construct. This report of an active plant Tc1/mariner in yeast will provide a foundation for future comparative analyses of animal and plant elements in addition to making a new wide host range transposable element available for plant gene tagging.
Tc1/水手转座因子超家族广泛分布于动植物基因组中。然而,此前尚未鉴定出有活性的植物元件。基因组中一个名为Osmar5的水稻(Oryza sativa)Tc1/水手元件的近乎相同的拷贝暗示了其潜在活性。先前的研究表明,Osmar5编码一种能特异性结合其自身末端的蛋白质。在本报告中,我们通过证明其编码序列在酵母中的表达促进了位于报告构建体中的非自主Osmar5元件的切除,表明Osmar5是一种活性转座因子。元件切除会产生转座子足迹,而元件重新插入则发生在与切除位点紧密相连或不相连的TA二核苷酸处。转座酶中的几个定点突变消除了活性,而转座酶结合位点的突变则阻止了非自主元件从报告构建体中转座。本报告中关于酵母中一种活性植物Tc1/水手元件的研究,将为未来动植物元件的比较分析奠定基础,此外还将提供一种新的广泛宿主范围的转座因子用于植物基因标签。