Wilson S, Matyunina L V, McDonald J F
Department of Genetics, University of Georgia, Athens, GA 30602, USA.
Gene. 1998 Mar 16;209(1-2):239-46. doi: 10.1016/s0378-1119(98)00048-1.
The untranslated leader region (ULR) of the Drosophila LTR retrotransposon copia is known to be critical to the element's expression in a variety of species. Two copia ULR size variants are prevalent in natural populations. The more transcriptionally active full length variants contain within their ULRs two tandemly repeated copies of a 28-bp region of dyad symmetry with a sequence similarity to the core sequence of the SV40 enhancer. The region of dyad symmetry contains two inverted repeats of a 8-bp motif (TTGTGAAA) that occurs at three additional locations within the ULR. The less active ULR gap variants differ from full length variants in that they contain only one copy of the 28-bp sequence. We show that the full length copia ULR in either orientation but not the gap ULR can significantly enhance expression of a minimal hsp 70 promoter. We demonstrate by EMSA that the full length ULR, the gap ULR and the 28-bp sequence are each capable of binding the Drosophila CCAAT/enhancer binding protein (DmC/EBP) and another previously uncharacterized factor, copia binding factor-1 (CBF-1). Another Drosophila protein previously implicated in fat body specific expression of the alcohol dehydrogenase gene (Adh), the Box-B-binding factor-2 (BBF-2), is also shown to bind to the copia ULR.
果蝇LTR逆转录转座子copia的未翻译前导区(ULR)已知对该元件在多种物种中的表达至关重要。两种copia ULR大小变体在自然种群中普遍存在。转录活性更高的全长变体在其ULR内包含两个28 bp的二元对称区域的串联重复拷贝,其序列与SV40增强子的核心序列相似。二元对称区域包含一个8 bp基序(TTGTGAAA)的两个反向重复,该基序在ULR内的另外三个位置出现。活性较低的ULR缺口变体与全长变体的不同之处在于它们仅包含一个28 bp序列的拷贝。我们表明,任一方向的全长copia ULR而非缺口ULR都能显著增强最小hsp 70启动子的表达。我们通过电泳迁移率变动分析(EMSA)证明,全长ULR、缺口ULR和28 bp序列均能够结合果蝇CCAAT/增强子结合蛋白(DmC/EBP)和另一种先前未鉴定的因子,即copia结合因子-1(CBF-1)。另一种先前与酒精脱氢酶基因(Adh)的脂肪体特异性表达有关的果蝇蛋白,即盒B结合因子-2(BBF-2),也被证明能与copia ULR结合。