A Three-protein-DNA Complex on a B Cell-specific Domain of the Immunoglobulin μ Heavy Chain Gene Enhancer*

A Three-protein-DNA Complex on a B Cell-specific Domain of the Immunoglobulin μ Heavy Chain Gene Enhancer*
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免疫球蛋白μ重链基因增强剂 B 细胞特异性结构域上的三蛋白 DNA 复合物*

DOI:
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发表时间:
1997
影响因子:
4.8
通讯作者:
R. Sen
R. Sen
中科院分区:
生物学2区
文献类型:
--
作者:
Elena Rao;W. Dang;Gang Tian;R. Sen

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淋巴细胞特异性免疫球蛋白μ重链基因内含子增强子(μ e)含有多个反式核因子的结合位点。我们使用了体外和体内实验的组合来重建一个最小的B细胞特异性μ增强子上的蛋白质- dna相互作用,该增强子包含三个基序,μ a, μB和μ e3。利用ets结构域蛋白在非淋巴样细胞中反激活最小增强子,我们发现(i)在体外,pu -1与μA和μB位点协调结合;(ii)在Ets-1存在的情况下,该因子与μA位点结合,pu -1与μB位点结合。两个因子TFE3和USF与μE3元素结合。当ETS蛋白与μE3结合蛋白一起存在时,产生一个三蛋白- dna复合物。此外,我们还提供了结合μA和μB位点的Ets-1和PU.1蛋白之间,以及结合μA和μE3位点的Ets-1和TFE3之间蛋白-蛋白相互作用的证据。我们提出μ增强子的这个结构域被组装成一个核蛋白复合体,该复合体包含两个组织限制性ETS结构域蛋白,它们识别来自螺旋同一侧的DNA,以及一个在它们之间结合的普遍表达的bhlh -亮氨酸拉链蛋白,它识别来自螺旋不同一侧的DNA。
The lymphoid-specific immunoglobulin μ heavy chain gene intron enhancer (μE) contains multiple binding sites for trans-acting nuclear factors. We have used a combination of in vitro and in vivo assays to reconstruct protein-DNA interactions on a minimal B cell-specific μ enhancer that contains three motifs, μA, μB, and μE3. Using ETS-domain proteins that transactivate the minimal enhancer in non-lymphoid cells, we show that (i) PU.1 binds coordinately to both μA and μB sites in vitro and (ii) in the presence of Ets-1, this factor binds to the μA site and PU.1 to the μB site. Two factors, TFE3 and USF, bind to the μE3 element. When the ETS proteins are present together with μE3 binding proteins, a three-protein-DNA complex is generated. Furthermore, we provide evidence for protein-protein interactions between Ets-1 and PU.1 proteins that bind to μA and μB sites, and between Ets-1 and TFE3 bound to the μA and μE3 sites. We propose that this domain of the μ enhancer is assembled into a nucleoprotein complex that contains two tissue-restricted ETS domain proteins that recognize DNA from the same side of the helix and one ubiquitously expressed bHLH-leucine zipper protein that binds between them, recognizing its site from a different side of the helix.
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