Fattash Isam, Lee Chia-Ni, Mo Kaiguo, Yang Guojun
Department of Biology, University of Toronto Mississauga, ON, Canada.
FEBS J. 2015 May;282(10):1829-40. doi: 10.1111/febs.13257. Epub 2015 Apr 8.
Miniature inverted repeat transposable elements (MITEs) are often the most numerous DNA transposons in plant and animal genomes. The dramatic amplification of MITE families during evolution is puzzling, because the transposase sources for the vast majority of MITE families are unknown. The yellow fever mosquito genome contains > 220-Mb MITE sequences; however, transposition activity has not been demonstrated for any of the MITE families. The Gnome elements are the youngest MITE family in this genome, with at least 116 identical copies. To test whether the putative autonomous element Ozma is capable of mobilizing Gnome and its two sibling MITEs, analyses were performed in a yeast transposition assay system. Whereas the wild-type transposase resulted in very low transposition activity, mutations in the region containing a putative nuclear export signal motif resulted in a dramatic (at least 4160-fold) increase in transposition frequency. We have also demonstrated that each residue of the novel DD37E motif is required for the activity of the Ozma transposase. Footprint sequences left at the donor sites suggest that the transposase may cleave between the second and the third nucleotides from the 5' ends of the elements. The excised elements reinsert specifically at dinucleotide 'TA', ~ 55% of them in yeast genes. The elements described in this article could potentially be useful as genetic tools for genetic manipulation of mosquitoes.
微型反向重复转座元件(MITEs)通常是植物和动物基因组中数量最多的DNA转座子。在进化过程中,MITE家族的急剧扩增令人费解,因为绝大多数MITE家族的转座酶来源尚不清楚。黄热病蚊子基因组包含超过220兆碱基的MITE序列;然而,尚未证明任何MITE家族具有转座活性。Gnome元件是该基因组中最年轻的MITE家族,至少有116个相同拷贝。为了测试假定的自主元件Ozma是否能够动员Gnome及其两个姊妹MITE,在酵母转座分析系统中进行了分析。野生型转座酶导致的转座活性非常低,而包含假定核输出信号基序的区域发生突变会导致转座频率急剧增加(至少增加4160倍)。我们还证明了新型DD37E基序的每个残基都是Ozma转座酶活性所必需的。供体位点留下的足迹序列表明,转座酶可能在元件5'端的第二个和第三个核苷酸之间切割。切除的元件特异性地重新插入二核苷酸“TA”,其中约55%插入酵母基因中。本文所述的元件可能作为遗传工具用于蚊子的基因操作。