Cvekl A, Sax C M, Li X, McDermott J B, Piatigorsky J
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892-2730, USA.
Proc Natl Acad Sci U S A. 1995 May 9;92(10):4681-5. doi: 10.1073/pnas.92.10.4681.
The abundance of delta-crystallin in the chicken eye lens provides an advantageous marker for tissue-specific gene expression during cellular differentiation. The lens-specific expression of the delta 1-crystallin gene is governed by an enhancer in the third intron, which binds a positive (delta EF2) and negative (delta EF1) factor in its core region. Here we show by DNase I footprinting, electrophoretic mobility-shift assays, and cotransfection experiments with the delta 1-promoter/enhancer fused to the chloramphenicol acetyltransferase reporter gene that the delta 1-crystallin enhancer has two adjacent functional Pax-6 binding sites. We also demonstrate by DNase I footprinting that the delta EF1 site can bind the transcription factor USF, raising the possibility that USF may cooperate with Pax-6 in activation of the chicken delta 1- and alpha A-crystallin genes. These data, coupled with our recent demonstration that Pax-6 activates the alpha A-crystallin gene, suggest that Pax-6 may have been used extensively throughout evolution to recruit and express crystallin genes in the lens.
鸡眼晶状体中大量的δ-晶体蛋白为细胞分化过程中的组织特异性基因表达提供了一个有利的标记。δ1-晶体蛋白基因的晶状体特异性表达受第三个内含子中的一个增强子调控,该增强子在其核心区域结合一个正向(δEF2)和一个负向(δEF1)因子。在这里,我们通过DNA酶I足迹分析、电泳迁移率变动分析以及将δ1-启动子/增强子与氯霉素乙酰转移酶报告基因融合进行的共转染实验表明,δ1-晶体蛋白增强子有两个相邻的功能性Pax-6结合位点。我们还通过DNA酶I足迹分析证明,δEF1位点可以结合转录因子USF,这增加了USF可能与Pax-6协同激活鸡δ1-和αA-晶体蛋白基因的可能性。这些数据,再加上我们最近证明Pax-6激活αA-晶体蛋白基因,表明Pax-6可能在整个进化过程中被广泛用于在晶状体中招募和表达晶体蛋白基因。