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Analysis of new molecular mechanism in transcriptional regulation using Drosophila

果蝇转录调控新分子机制分析

基本信息

批准号：
18570161
负责人：
UEDA Hitoshi
金额：
$ 2.57万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Transgenic fly promoter assay revealed that several conserved sequences between Drosophila melanogaster and related spices and DHR3 binding site which locates 140 and 100 bp, respectively, from the transcriptional start site of the EDG84A gene are important for high level expression of the EDG84A gene. Furthermore, we showed that ecdysone-inducible transcriptional repressor Blimp-1 plays an important role to restrict the expression timing of the transcription factor FTZ-Fl which determines expression timing of the EDG84A gene. These findings and previous results indicate that temporal and spatial expression of the EDG84A gene is controlled by complicated mechanism including several transcription factors.Effects of insertion positions of transgenes carrying the EDG84A promoter-LacZ fusion gene or the EDG78E promoter- LacZ fusion gene in newly established transgenic fly lines were analyzed by detecting expression of the white gene indicated by eye color. In the fly carrying the EDG84A promoter-LacZ fusion gene, 85% of the lines showed the pairing dependent activation of the white gene and the rest of the 15% did not show it. On the other hand, only 18% of the established lines carrying the EDG78E promoter showed the pairing dependent activation of the white gene and 82% did not show it. These results suggest that (1) the EDG84A promoter sequence enhances the pairing dependent activation and the EDG78E promoter basically dose not have such effect, and (2) enhancing and suppressing regions for the pairing dependent gene activation are present within the genome in relatively high efficiency.

转基因果蝇启动子分析表明，果蝇与相关香料之间的几个保守序列以及EDG84A基因转录起始点上分别位于140和100bp的DHR3结合位点对EDG84A基因的高效表达是重要的。此外，我们还发现，蜕皮激素诱导的转录抑制因子Blimp-1在限制转录因子FTZ-F1的表达时机方面发挥了重要作用，FTZ-F1决定了EDG84A基因的表达时机。这些结果和以前的结果表明，EDG84A基因的时空表达受多个转录因素的复杂机制控制。通过检测眼睛颜色指示的白色基因的表达，分析了携带EDG84A启动子-LacZ融合基因或EDG78E启动子-LacZ融合基因的转基因在新建立的转基因果蝇系中的插入位置的影响。在携带EDG84A启动子-LacZ融合基因的果蝇中，85%的品系表现出白色基因的配对依赖激活，其余15%的品系没有表现出这种激活。另一方面，在已建立的携带EDG78E启动子的品系中，只有18%的品系表现出白色基因的配对依赖激活，82%的品系没有表现出这种激活。这些结果表明：(1)EDG84A启动子序列增强配对依赖激活，而EDG78E启动子基本没有这种作用；(2)基因组中存在相对高效的配对依赖基因激活增强和抑制区域。