Transcriptional regulation of the Drosophila orc2 gene by the DREF pathway
Transcriptional regulation of the Drosophila orc2 gene by the DREF pathway
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DOI：
10.1016/j.bbaexp.2005.10.009
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发表时间：
2005-12-30
期刊：
影响因子：
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通讯作者：
Yamaguchi, M
中科院分区：
文献类型：
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作者：
Okudaira, K;Ohno, K;Yamaguchi, M
DNA复制相关元件（DRE）和DREF结合因子（DREF）在果蝇中调控PCNA和DNA聚合酶α等DNA复制相关基因中起重要作用。我们之前报道过，在发育中的眼成像盘中，DREF过表达可诱导异位DNA合成和细胞凋亡，从而导致粗糙的眼睛。为了确定与DREF基因的遗传相互作用，我们对粗糙眼表型的修饰因子进行了筛选。其中一个被发现的抑制基因是果蝇orc2基因。通过对已知转录因子识别位点的搜索发现，orc2基因包含三个DREs，分别为DRE1（+14 ~ +21）、DRE2（-205 ~ -198）和DRE3（-709 ~ -702）。利用Kc细胞核提取物进行能带迁移分析，检测DREF与DRE1或DRE2之间形成的特定络合物。染色质免疫沉淀实验进一步证明了DREF与含有DRE1或DRE2的基因组区域的特异性结合，表明这些是体内形成的真正复合物。Kc细胞的荧光素酶测定表明，orc2启动子中的DRE位点参与了orc2基因的转录调控。结果表明，orc2基因受DREF通路的控制。(c) 2005 Elsevier B.V.版权所有
DNA replication-related element (DRE) and the DRE-binding factor (DREF) play an important role in regulating DNA replication-related genes such as PCNA and DNA polymerase alpha in Drosophila. We have previously reported that overexpression of DREF in developing eye imaginal discs induced ectopic DNA synthesis and apoptosis, which results in rough eyes. To identify genetic interactants with the DREF gene, we have carried out a screen for modifiers of the rough eye phenotype. One of the suppressor genes identified was the Drosophila orc2 gene. A search for known transcription factor recognition sites revealed that the orc2 gene contains three DREs, named DRE1 (+14 to +21), DRE2 (-205 to -198), and DRE3 (-709 to -702). Band mobility shift analysis using Kc cell nuclear extracts detected the specific complex formed between DREF and the DRE1 or DRE2. Specific binding of DREF to genomic region containing the DRE1 or DRE2 was further demonstrated by chromatin immunoprecipitation assays, suggesting that these are the genuine complexes formed in vivo. The luciferase assay in Kc cells indicated that the DRE sites in the orc2 promoter are involved in a transcriptional regulation of the orc2 gene. The results, taken together, demonstrate that the orc2 gene is under the control of DREF pathway. (c) 2005 Elsevier B.V. All rights reserved.