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果蝇 AP-2 转录因子同源物的克隆与特性分析

Cloning and characterization of the Drosophila homologue of the AP-2 transcription factor.

作者信息

Bauer R, McGuffin M E, Mattox W, Tainsky M A

机构信息

Department of Tumor Biology, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.

出版信息

Oncogene. 1998 Oct 15;17(15):1911-22. doi: 10.1038/sj.onc.1202114.

DOI:10.1038/sj.onc.1202114
PMID:9788434
Abstract

The AP-2 family of transcription factors (AP-2alpha, AP-2beta and AP-2gamma) is temporally and spatially regulated in mammals and has also been implicated in oncogenesis. Here we report the isolation of genomic and cDNA clones encoding the Drosophila homologue of AP-2, designated DAP-2. The predicted amino acid sequence exhibits 42-45% overall identity with the vertebrate AP-2 proteins. A sequence of 107 amino acids within the DNA binding and dimerization domain of the vertebrate AP-2 proteins is highly conserved (90-92%) with the DAP-2 homologue. An in vitro translation product of DAP-2 cDNA binds specifically to AP-2 consensus binding sites. DAP-2 was also shown to be functionally conserved in vivo because transient transfection of a DAP-2 expression plasmid activated transcription through AP-2 binding sites in both mammalian and Drosophila cell lines. DAP-2 is expressed during early embryogenesis and DAP-2 transcripts are also detected in the adult. Whole-mount in situ hybridizations demonstrated that DAP-2 is expressed initially at stage 9 of Drosophila embryonic development and that DAP-2 transcripts are detected in regions of the brain, eye-antennal disc, optical lobe, antenno-maxillary complex, and in a subset of cells of the ventral nerve cord. The cloning of DAP-2 and the identification of the DAP-2 expression pattern during embryogenesis provides a starting point to address the function of AP-2 during differentiation and development in a well understood model system.

摘要

转录因子AP-2家族(AP-2α、AP-2β和AP-2γ)在哺乳动物中受到时空调控,并且也与肿瘤发生有关。在此,我们报告了果蝇AP-2同源物(命名为DAP-2)的基因组和cDNA克隆的分离。预测的氨基酸序列与脊椎动物AP-2蛋白的总体一致性为42%-45%。脊椎动物AP-2蛋白的DNA结合和二聚化结构域内的107个氨基酸序列与DAP-2同源物高度保守(90%-92%)。DAP-2 cDNA的体外翻译产物特异性结合AP-2共有结合位点。DAP-2在体内也表现出功能保守性,因为DAP-2表达质粒的瞬时转染激活了哺乳动物和果蝇细胞系中通过AP-2结合位点的转录。DAP-2在胚胎发育早期表达,在成体中也可检测到DAP-2转录本。全胚胎原位杂交表明,DAP-2最初在果蝇胚胎发育的第9阶段表达,并且在脑、眼触角盘、视叶、触角-上颌复合体以及腹神经索的一部分细胞区域中检测到DAP-2转录本。DAP-2的克隆以及胚胎发育过程中DAP-2表达模式的鉴定为在一个易于理解的模型系统中研究AP-2在分化和发育过程中的功能提供了一个起点。

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