Fetal Alz-50 clone 1, a novel zinc finger protein, binds a specific DNA sequence and acts as a transcriptional regulator
Fetal Alz-50 clone 1, a novel zinc finger protein, binds a specific DNA sequence and acts as a transcriptional regulator
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DOI：
10.1074/jbc.274.49.35262
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发表时间：
1999-12-03
影响因子：
4.8
通讯作者：
Bowser, R
中科院分区：
文献类型：
--
作者：
Jordan-Sciutto, KL;Dragich, JM;Bowser, R
胎儿Alz-50克隆1 （FAC1）是一种新的发育调控基因，在神经元发育和神经退行性变过程中表现出蛋白质表达和亚细胞定位的变化。为了了解亚细胞定位改变的功能含义，我们建立了FAC1的正常细胞功能，FAC1氨基酸序列与转录调控因子具有区域同源性。利用聚合酶链反应辅助的结合位点选择试验，我们鉴定了重组FAC1识别的DNA序列，鉴定的结合位点上任意两个相邻碱基对的突变显著降低了FAC1的结合偏好，表明该结合对鉴定的位点具有特异性。神经和非神经细胞系的核提取物含有与重组FAC1蛋白相似的特异性和核苷酸要求的dna结合活性，这种dna结合活性可归因于FAC1，因为它依赖于FAC1的存在，并且在非变性聚丙烯酰胺凝胶上的行为与瞬时转染的FAC1相同。在NIH3T3细胞中，含有鉴定结合位点（CACAACAC）的荧光素酶报告质粒被共转染的FAC1抑制，无论结合位点在转录起始位点的近端还是远端。本研究表明，FAC1是一种dna结合蛋白，当定位于细胞核时作为转录因子发挥作用。
Fetal Alz-50 clone 1 (FAC1) is a novel, developmentally regulated gene that exhibits changes in protein expression and subcellular localization during neuronal development and neurodegeneration, To understand the functional implications of altered subcellular localization, we have established a normal cellular function of FAC1, The FAC1 amino acid sequence contains regional homology to transcriptional regulators. Using the polymerase chain reaction-assisted binding site selection assay, we have identified a DNA sequence recognized by recombinant FAC1, Mutation of any 2 adjacent base pairs in the identified binding site dramatically reduced the binding preference of FAC1, demonstrating that the binding is specific for the identified site. Nuclear extracts from neural and non-neural cell lines contained a DNA-binding activity with similar specificity and nucleotide requirements as the recombinant FAC1 protein, This DNA-binding activity can be attributed to FAC1 since it is dependent upon the presence of FAC1 and behaves identically on a nondenaturing polyacrylamide gel as transiently transfected FAC1. In NIH3T3 cells, luciferase reporter plasmids containing the identified binding site (CACAACAC) were repressed by cotransfected FAC1 whether the binding site was proximal or distal to the transcription initiation site. This study indicates that FAC1 is a DNA-binding protein that functions as a transcription factor when localized to the nucleus.