Characterization of dRFX2, a novel RFX family protein in Drosophila
Characterization of dRFX2, a novel RFX family protein in Drosophila
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DOI：
10.1093/nar/gkh895
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Yamaguchi, M
中科院分区：
文献类型：
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作者：
Otsuki, K;Hayashi, Y;Yamaguchi, M
在果蝇PCNA基因启动子上游调控元件（URE）和DNA复制相关元件（DRE）之间的区域中发现了一个转录调控元件。该元件在活蝇的启动子活性中起重要作用。使用该元件作为诱饵的酵母单杂交筛选允许分离编码在体外与该元件结合的蛋白质的cDNA。核苷酸序列分析表明，该cDNA编码一种新的蛋白质，含有一个特征性的DNA结合结构域之间的调节因子X（RFX）家族蛋白质的保守。我们将该蛋白质命名为果蝇RFX 2（dRFX 2），并将该元件命名为dRFX 2位点。为了研究dRFX 2在体内的功能，我们采取了分析对内源性dRFX 2的显性负效应的策略。建立转基因果蝇，其中携带含有dRFX 2的RFX结构域（DNA结合结构域）的202至480的氨基酸序列的HA-dRFX（202-480）的表达靶向于眼睛成虫盘中的细胞。在表达HA-dRFX（202-480）的眼视盘中，尽管感光细胞正常分化，但G1-S转换和/或S期进展被中断，并诱导异位凋亡。这些结果表明dRFX 2在G1-S转变和/或S期进展中起作用。
A transcriptional regulatory element was identified in the region between URE (upstream regulatory element) and DRE (DNA replication-related element) in the Drosophila PCNA gene promoter. This element plays an important role in promoter activity in living flies. A yeast one-hybrid screening using this element as a bait allowed isolation of a cDNA encoding a protein which binds to the element in vitro. Nucleotide sequence analyses revealed that the cDNA encodes a novel protein containing a characteristic DNA-binding domain conserved among the regulatory factor X (RFX) family proteins. We termed this protein Drosophila RFX2 (dRFX2) and this element dRFX2 site. To investigate the function of dRFX2 in vivo, we took the strategy of analyzing the dominant negative effects against the endogenous dRFX2. Transgenic flies were established in which expression of HA-dRFX(202-480) carrying the amino acid sequences from 202 to 480 containing the RFX domain (DNA-binding domain) of dRFX2 was targeted to the cells in the eye imaginal discs. In the eye imaginal disc expressing the HA-dRFX(202-480), the G1-S transition and/or the progression of S phase were/was interrupted, and the ectopic apoptosis was induced, though photoreceptor cells differentiated normally. These results indicate that dRFX2 plays a role in G1-S transition and/or in progression of S phase.