DeSimone S M, White K
Department of Biology, Brandeis University, Waltham, Massachusetts 02254.
Mol Cell Biol. 1993 Jun;13(6):3641-9. doi: 10.1128/mcb.13.6.3641-3649.1993.
The erect wing (ewg) locus of Drosophila melanogaster encodes a vital function important for the development of the nervous system and the indirect flight muscles. In order to understand the ewg function at a molecular level, cDNA clones were isolated. Sequence analysis of cDNAs revealed a single open reading frame (ORF) encoding a protein of 733 residues. The translational start for this ORF is a CTG codon. A 225-amino-acid region of this protein is 71% identical to the DNA binding region of the Strongylocentrotus purpuratus P3A2 DNA binding protein. Additionally, the ORF contains large acidic and basic domains characteristic of those in proteins involved in nuclear regulatory functions. Immunoblot analysis using polyclonal anti-EWG antisera generated against a bacterial fusion protein reveals a single, 116-kDa protein present throughout development, beginning at approximately stage 12 of embryogenesis, which is enriched in adult heads and absent from embryos carrying certain ewg alleles. Additionally, we show that EWG is localized specifically to the nuclei of virtually all embryonic neurons. Finally, a minigene consisting of an ewg cDNA under control of the hsp70 promoter can provide the ewg function in transgenic ewg mutant flies.
黑腹果蝇的直立翅(ewg)基因座编码一种对神经系统和间接飞行肌发育至关重要的功能。为了在分子水平上理解ewg的功能,分离了cDNA克隆。对cDNA的序列分析揭示了一个单一的开放阅读框(ORF),其编码一个由733个残基组成的蛋白质。该ORF的翻译起始密码子是CTG。该蛋白质的一个225个氨基酸的区域与紫海胆P3A2 DNA结合蛋白的DNA结合区域有71%的同一性。此外,该ORF包含与参与核调节功能的蛋白质中常见的大酸性和碱性结构域。使用针对细菌融合蛋白产生的多克隆抗EWG抗血清进行的免疫印迹分析显示,在整个发育过程中存在一种单一的、116 kDa的蛋白质,从胚胎发育的大约第12阶段开始出现,在成年头部富集,而在携带某些ewg等位基因的胚胎中不存在。此外,我们表明EWG特异性地定位于几乎所有胚胎神经元的细胞核中。最后,一个由hsp70启动子控制的ewg cDNA组成的微型基因可以在转基因ewg突变果蝇中提供ewg功能。