Hirochika H, Otsuki H, Yoshikawa M, Otsuki Y, Sugimoto K, Takeda S
Department of Molecular Biology, National Institute of Agrobiological Resources, Ibaraki, Japan.
Plant Cell. 1996 Apr;8(4):725-34. doi: 10.1105/tpc.8.4.725.
The complete nucleotide sequence of the tobacco retrotransposon Tto1, one of the few active retrotransposons of plants, was determined. The sequence analysis suggests that Tto1 carries all functions required for autonomous transposition through reverse transcription. Gene organization and the nature of the transcription product suggest that Tto1 uses a gene expression mechanism different from those employed by retroviruses and most retrotransposons to regulate Gag and Pol stoichiometry. Tto1 was introduced into rice to study its autonomous transposition in heterologous hosts. Transcription and transposition of Tto1 were observed in rice cells. To probe the autonomous transposition through reverse transcription, a modified Tto1 retrotransposon in which part of a reverse transcriptase gene was replaced with an intron-containing hygromycin resistance gene was constructed and introduced into rice cells. Loss of the intron was observed only when intact Tto1 was cotransfected. These results indicate that Tto1 can transpose autonomously through reverse transcription and that the host factors required for transposition are conserved among monocots (class Magnoliopsida; rice) and dicots (class Liliopsida; tobacco), which diverged approximately 200 million years ago. These findings are discussed in relation to the regulation and evolution of retrotransposons and the possible use of Tto1 as a molecular genetic tool.
烟草逆转录转座子Tto1是植物中少数活跃的逆转录转座子之一,其完整的核苷酸序列已被测定。序列分析表明,Tto1具备通过逆转录进行自主转座所需的所有功能。基因组织和转录产物的性质表明,Tto1使用一种不同于逆转录病毒和大多数逆转录转座子所采用的基因表达机制来调节Gag和Pol的化学计量。将Tto1导入水稻以研究其在异源宿主中的自主转座。在水稻细胞中观察到了Tto1的转录和转座。为了探究通过逆转录进行的自主转座,构建了一种修饰的Tto1逆转录转座子,其中逆转录酶基因的一部分被含内含子的潮霉素抗性基因所取代,并将其导入水稻细胞。仅当完整的Tto1共转染时才观察到内含子的缺失。这些结果表明,Tto1可以通过逆转录进行自主转座,并且转座所需的宿主因子在大约2亿年前分化的单子叶植物(木兰纲;水稻)和双子叶植物(百合纲;烟草)中是保守的。结合逆转录转座子的调控和进化以及Tto1作为分子遗传工具的可能用途对这些发现进行了讨论。