Drummond I A, Rohwer-Nutter P, Sukhatme V P
Harvard Medical School, Boston, MA.
DNA Cell Biol. 1994 Oct;13(10):1047-55. doi: 10.1089/dna.1994.13.1047.
The Egr family of transcriptional regulators comprises a group of genes which encode members of the Cys2-His2 class of zinc-finger proteins. We have isolated a zebrafish egr1 homologue by screening a zebrafish genomic library with a mouse Egr1 zinc finger probe. Southern blotting indicated the existence of a single zebrafish egr1 gene and, as in higher vertebrates, the presence of related members of a larger gene family. Sequence analysis of the zebrafish egr1 coding region revealed a high level of homology to the mouse, rat, and human Egr1 genes with the notable exception of a polymorphic, triplet nucleotide repeat sequence in the region coding for the amino terminus of the Egr1 protein. The predicted DNA-binding, zinc-finger domain protein sequence was strictly conserved. The 5' region of the zebrafish egr1 gene contained a variety of transcription factor binding sites, also present in the mouse gene, for serum response factor, CREB and c-Ets. The zebrafish egr1 transcript was approximately 3.4 kb in size and was expressed in adult zebrafish brain and muscle RNA, a pattern of expression similar to that observed in mice. The potential for zebrafish egr1 to function as a transcriptional regulator was tested by constructing an expression vector containing zebrafish egr1 coding sequences under the control of a cytomegalovirus promoter. This construct was found to activate transcription of a reporter plasmid bearing multiple Egr1 binding sites when transiently cotransfected into mouse 3T3 cells. Our results indicate that the structure, regulation, and function of the Egr1 gene have been highly conserved during vertebrate evolution and suggest an important role for this gene in growth and development.
转录调节因子Egr家族由一组基因组成,这些基因编码Cys2-His2类锌指蛋白成员。我们用小鼠Egr1锌指探针筛选斑马鱼基因组文库,分离出了斑马鱼egr1同源物。Southern印迹分析表明斑马鱼中存在单个egr1基因,并且与高等脊椎动物一样,存在一个更大基因家族的相关成员。斑马鱼egr1编码区的序列分析显示,它与小鼠、大鼠和人类的Egr1基因具有高度同源性,但在编码Egr1蛋白氨基末端的区域存在一个多态性三联体核苷酸重复序列,这是一个显著的例外。预测的DNA结合锌指结构域蛋白序列严格保守。斑马鱼egr1基因的5'区域包含多种转录因子结合位点,小鼠基因中也存在这些位点,用于血清反应因子、CREB和c-Ets。斑马鱼egr1转录本大小约为3.4 kb,在成年斑马鱼的脑和肌肉RNA中表达,这种表达模式与在小鼠中观察到的相似。通过构建一个在巨细胞病毒启动子控制下包含斑马鱼egr1编码序列的表达载体,测试了斑马鱼egr1作为转录调节因子的功能。当将该构建体与报告质粒瞬时共转染到小鼠3T3细胞中时,发现它能激活带有多个Egr1结合位点的报告质粒的转录。我们的结果表明,Egr1基因的结构、调节和功能在脊椎动物进化过程中高度保守,并表明该基因在生长和发育中起重要作用。