Liu D, Wang R, Galli M, Crawford N M
Section of Cell and Developmental Biology, Division of Biology, University of California at San Diego, La Jolla, California 92093-0116, USA.
Plant Cell. 2001 Aug;13(8):1851-63. doi: 10.1105/tpc.010030.
Various sequences within Tag1, the endogenous transposon of Arabidopsis, were examined to determine how Tag1 excision and expression are regulated. The 5' intron for the major 2.3-kb Tag1 transcript was found to be critical for the accumulation of Tag1 transcripts and for high rates of somatic excision. This was true for the autonomous element in cauliflower mosaic virus 35S-Tag1-beta-glucuronidase constructs and for a two-component system using the 35S promoter to produce Tag1 transposase and a beta-glucuronidase::dTag1 marker construct to score for excision. The 3' introns of Tag1, although not needed for high transposase expression in primary transgenic plants, were important for maintaining high levels of somatic excision and accumulation of the major but not the minor Tag1 transcripts in subsequent generations. With both 5' and 3' introns present, exchanging the 5' promoter region of Tag1 with the 35S promoter did not affect the timing of Tag1 excision significantly, but it did disrupt germinal excision. Removal of the 5' intron did not abolish germinal excision activity, however. These results indicate that somatic and germinal excision of Tag1 are differentially controlled, with the 5' promoter region being critical for germinal excision activity and the 5' intron playing an important role for somatic excision, possibly via intron-mediated enhancement.
对拟南芥内源性转座子Tag1中的各种序列进行了研究,以确定Tag1的切除和表达是如何调控的。发现主要的2.3kb Tag1转录本的5'内含子对于Tag1转录本的积累和高频率的体细胞切除至关重要。对于花椰菜花叶病毒35S-Tag1-β-葡萄糖醛酸酶构建体中的自主元件以及使用35S启动子产生Tag1转座酶和β-葡萄糖醛酸酶::dTag1标记构建体来计分切除的双组分系统而言,情况都是如此。Tag1的3'内含子虽然在初级转基因植物中对于高转座酶表达不是必需的,但对于在后代中维持高水平的体细胞切除和主要而非次要Tag1转录本的积累很重要。当同时存在5'和3'内含子时,将Tag1的5'启动子区域与35S启动子交换不会显著影响Tag1切除的时间,但会破坏生殖细胞切除。然而,去除5'内含子并没有消除生殖细胞切除活性。这些结果表明,Tag1的体细胞切除和生殖细胞切除受到不同的控制,5'启动子区域对于生殖细胞切除活性至关重要,而5'内含子可能通过内含子介导的增强作用在体细胞切除中发挥重要作用。