Amero S A, Elgin S C, Beyer A L
Department of Microbiology, University of Virginia School of Medicine, Charlottesville 22908.
Genes Dev. 1991 Feb;5(2):188-200. doi: 10.1101/gad.5.2.188.
Using an immunochemical approach, we have identified a unique antigen, PEP (protein on ecdysone puffs), which is associated in third-instar larvae with the active ecdysone-regulated loci on polytene chromosomes; PEP is not associated with most intermolt puffs and is found on some, but not all, heat shock-induced puffs. The distribution pattern changes with changing puffing patterns in the developmental program. We have screened an expression library and recovered a cDNA clone encoding PEP. PEP possesses multiple potential nucleic acid- and protein- binding regions: a glycine- and asparagine-rich amino terminus, four zinc finger motifs, two very acidic segments, two short basic stretches, and an alanine- and proline-rich carboxyl terminus. The Pep gene maps by in situ hybridization to the cytological locus 74F, adjacent to the early ecdysone-responsive region; however, the gene is not regulated by ecdysone at the level of transcription. The pattern of Pep expression through development suggests that maternal Pep gene transcripts are supplied to the embryo, and that the abundance of Pep gene transcripts decreases to a lower, fairly constant level thereafter. This unusual protein may play a role in the process of gene activation, or possibly in RNA processing, for a defined set of developmentally regulated loci.
利用免疫化学方法,我们鉴定出一种独特的抗原,即蜕皮激素胀泡蛋白(PEP),它在三龄幼虫中与多线染色体上活跃的蜕皮激素调控位点相关联;PEP与大多数蜕皮间期胀泡不相关,且在一些但并非所有热激诱导的胀泡上存在。其分布模式随发育程序中胀泡模式的变化而改变。我们筛选了一个表达文库并获得了一个编码PEP的cDNA克隆。PEP具有多个潜在的核酸和蛋白质结合区域:富含甘氨酸和天冬酰胺的氨基末端、四个锌指基序、两个非常酸性的区段、两个短的碱性片段以及富含丙氨酸和脯氨酸的羧基末端。通过原位杂交,Pep基因定位于细胞学位点74F,与早期蜕皮激素反应区域相邻;然而,该基因在转录水平不受蜕皮激素调控。Pep在整个发育过程中的表达模式表明,母体Pep基因转录本被提供给胚胎,此后Pep基因转录本的丰度降至较低且相当恒定的水平。这种不寻常的蛋白质可能在一组特定的发育调控位点的基因激活过程中发挥作用,或者可能在RNA加工过程中发挥作用。