Galli Mary, Theriault Angie, Liu Dong, Crawford Nigel M
Section of Cell and Developmental Biology, Division of Biological Sciences, University of California, San Diego, California 92093-0116, USA.
Genetics. 2003 Dec;165(4):2093-105. doi: 10.1093/genetics/165.4.2093.
The Arabidopsis transposon Tag1 undergoes late excision during vegetative and germinal development in plants containing 35S-Tag1-GUS constructs. To determine if transcriptional regulation can account for the developmental control of Tag1 excision, the transcriptional activity of Tag1 promoter-GUS fusion constructs of various lengths was examined in transgenic plants. All constructs showed expression in the reproductive organs of developing flowers but no expression in leaves. Expression was restricted to developing gametophytes in both male and female lineages. Quantitative RT-PCR analysis confirmed that Tag1 expression predominates in the reproductive organs of flower buds. These results are consistent with late germinal excision of Tag1, but they cannot explain the vegetative excision activity of Tag1 observed with 35S-Tag1-GUS constructs. To resolve this issue, Tag1 excision was reexamined using elements with no adjacent 35S promoter sequences. Tag1 excision in this context is restricted to germinal events with no detectable vegetative excision. If a 35S enhancer sequence is placed next to Tag1, vegetative excision is restored. These results indicate that the intrinsic activity of Tag1 is restricted to germinal excision due to targeted expression of the Tag1 transposase to developing gametophytes and that this activity is altered by the presence of adjacent enhancers or promoters.
在含有35S-Tag1-GUS构建体的植物中,拟南芥转座子Tag1在营养和生殖发育过程中经历晚期切除。为了确定转录调控是否可以解释Tag1切除的发育控制,在转基因植物中检测了各种长度的Tag1启动子-GUS融合构建体的转录活性。所有构建体在发育中的花的生殖器官中均有表达,但在叶片中无表达。表达仅限于发育中的雄配子体和雌配子体。定量RT-PCR分析证实,Tag1在花芽的生殖器官中表达占主导。这些结果与Tag1的晚期生殖切除一致,但无法解释用35S-Tag1-GUS构建体观察到的Tag1的营养切除活性。为了解决这个问题,使用没有相邻35S启动子序列的元件重新检查了Tag1切除。在这种情况下,Tag1切除仅限于生殖事件,没有可检测到的营养切除。如果将35S增强子序列置于Tag1旁边,则营养切除得以恢复。这些结果表明,由于Tag1转座酶靶向表达于发育中的配子体,Tag1的内在活性仅限于生殖切除,并且这种活性会因相邻增强子或启动子的存在而改变。