Nuclear factors binding to the extensin promoter exhibit differential activity in carrot protoplasts and cells.
Nuclear factors binding to the extensin promoter exhibit differential activity in carrot protoplasts and cells.
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与延伸蛋白启动子结合的核因子在胡萝卜原生质体和细胞中表现出不同的活性。
DOI：
10.1007/bf00020015
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发表时间：
1992
影响因子：
5.1
通讯作者：
Cashmore,AR
中科院分区：
文献类型：
--
作者：
Granell,A;Peretó,JG;Schindler,U;Cashmore,AR
细胞壁蛋白延伸蛋白（一种富含羟基脯氨酸的糖蛋白）的表达可由几种不同的刺激（包括损伤）诱导。原生质体的制备过程模拟了损伤效应，导致了伸展蛋白的产生。利用原生质体中的瞬时表达，我们分析了扩展蛋白启动子的几个缺失。我们发现了一个重要的转录调控元件，位于两个表征扩展蛋白启动子的TATA盒之间。其他调控元件位于- 719至- 658之间的上游，是最高表达水平所必需的。利用电泳迁移率转移测定和甲基化干扰实验，我们证明了核因子与这些上游调控元件的相互作用。除了先前发现的主要存在于未损伤细胞中的因子EGBF-1和EGBF-2外，我们还发现了一种新的dna结合活性，这种活性存在于原生质体制备的提取物中，而不存在于未损伤细胞的提取物中。这个因子被称为EBF（延伸蛋白结合蛋白），它与DNA片段结合，当删除该片段时，其表达会减少48%。
The expression of the cell wall protein extensin, a hydroxyproline-rich glycoprotein, is induced by several different stimuli, including wounding. The process of protoplast preparation mimics the wounding effect and results in the induction of extensin. Using transient expression in protoplasts we analyzed several deletions of the extensin promoter. We identified an important transcriptional regulatory element located between the two TATA boxes that characterize the extensin promoter. Other regulatory elements, located further upstream between −719 to −658, are necessary for maximum level of expression. Employing electrophoretic mobility shift assays and methylation interference experiments, we demonstrate the interaction of nuclear factors with these upstream regulatory elements. In addition to the previously identified factors EGBF-1 and EGBF-2, which are mainly present in unwounded cells, we identified an additional novel DNA-binding activity that is present in extracts prepared from protoplasts but not in extracts from unwounded cells. This factor, designated EBF (extensin-binding protein), binds to a DNA fragment which when deleted results in a 48% reduction in expression.