Zhao Dongyan, Ferguson Ann, Jiang Ning
Department of Horticulture, Michigan State University, East Lansing, Michigan 48824.
Department of Horticulture, Michigan State University, East Lansing, Michigan 48824
Plant Cell. 2015 Jan;27(1):132-48. doi: 10.1105/tpc.114.128488. Epub 2015 Jan 13.
Mutator-like transposable elements (MULEs) are widespread in plants and are well known for their high transposition activity as well as their ability to duplicate and amplify host gene fragments. Despite their abundance and importance, few active MULEs have been identified. In this study, we demonstrated that a rice (Oryza sativa) MULE, Os3378, is capable of excising and reinserting in yeast (Saccharomyces cerevisiae), suggesting that yeast harbors all the host factors for the transposition of MULEs. The transposition activity induced by the wild-type transposase is low but can be altered by modification of the transposase sequence, including deletion, fusion, and substitution. Particularly, fusion of a fluorescent protein to the transposase enhanced the transposition activity, representing another approach to manipulate transposases. Moreover, we identified a critical region in the transposase where the net charge of the amino acids seems to be important for activity. Finally, transposition efficiency is also influenced by the element and its flanking sequences (i.e., small elements are more competent than their large counterparts). Perfect target site duplication is favorable, but not required, for precise excision. In addition to the potential application in functional genomics, this study provides the foundation for further studies of the transposition mechanism of MULEs.
类Mutator转座元件(MULEs)在植物中广泛存在,以其高转座活性以及复制和扩增宿主基因片段的能力而闻名。尽管它们数量众多且很重要,但已鉴定出的活性MULEs却很少。在本研究中,我们证明了水稻(Oryza sativa)的一个MULE,即Os3378,能够在酵母(Saccharomyces cerevisiae)中切除并重新插入,这表明酵母中存在MULEs转座所需的所有宿主因子。野生型转座酶诱导的转座活性较低,但可以通过对转座酶序列进行修饰来改变,包括缺失、融合和替换。特别是,将荧光蛋白与转座酶融合可增强转座活性,这代表了另一种操纵转座酶的方法。此外,我们在转座酶中鉴定出一个关键区域,其中氨基酸的净电荷似乎对活性很重要。最后,转座效率也受元件及其侧翼序列的影响(即小元件比大元件更具转座能力)。完美的靶位点重复有利于精确切除,但不是必需的。除了在功能基因组学中的潜在应用外,本研究为进一步研究MULEs的转座机制奠定了基础。