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首页> 外文期刊>Molecular and Cellular Biology >Cloning of a negative transcription factor that binds to the upstream conserved region of Moloney murine leukemia virus.
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Cloning of a negative transcription factor that binds to the upstream conserved region of Moloney murine leukemia virus.

机译：克隆与莫洛尼鼠白血病病毒上游保守区结合的负转录因子。

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The long terminal repeat of Moloney murine leukemia virus (MuLV) contains the upstream conserved region (UCR). The UCR core sequence, CGCCATTTT, binds a ubiquitous nuclear factor and mediates negative regulation of MuLV promoter activity. We have isolated murine cDNA clones encoding a protein, referred to as UCRBP, that binds specifically to the UCR core sequence. Gel mobility shift assays demonstrate that the UCRBP fusion protein expressed in bacteria binds the UCR core with specificity identical to that of the UCR-binding factor in the nucleus of murine and human cells. Analysis of full-length UCRBP cDNA reveals that it has a putative zinc finger domain composed of four C2H2 zinc fingers of the GLI subgroup and an N-terminal region containing alternating charges, including a stretch of 12 histidine residues. The 2.4-kb UCRBP message is expressed in all cell lines examined (teratocarcinoma, B- and T-cell, macrophage, fibroblast, and myocyte), consistent with the ubiquitous expression of the UCR-binding factor. Transient transfection of an expressible UCRBP cDNA into fibroblasts results in down-regulation of MuLV promoter activity, in agreement with previous functional analysis of the UCR. Recently three groups have independently isolated human and mouse UCRBP. These studies show that UCRBP binds to various target motifs that are distinct from the UCR motif: the adeno-associated virus P5 promoter and elements in the immunoglobulin light- and heavy-chain genes, as well as elements in ribosomal protein genes. These results indicate that UCRBP has unusually diverse DNA-binding specificity and as such is likely to regulate expression of many different genes.
机译：莫洛尼氏鼠白血病病毒（MuLV）的长末端重复序列包含上游保守区（UCR）。 UCR核心序列CGCCATTTT与普遍存在的核因子结合，并介导MuLV启动子活性的负调控。我们已经分离出了鼠类cDNA克隆，该克隆编码一种称为UCRBP的蛋白，该蛋白与UCR核心序列特异性结合。凝胶迁移率变动分析表明，细菌中表达的UCRBP融合蛋白以与鼠和人细胞核中UCR结合因子相同的特异性结合UCR核心。全长UCRBP cDNA的分析表明，它具有一个推定的锌指结构域，该结构由四个GLI亚组的C2H2锌指和一个N端区域组成，该区域包含交替电荷，包括一段12个组氨酸残基。在所有检查的细胞系（畸胎癌，B细胞和T细胞，巨噬细胞，成纤维细胞和肌细胞）中表达了2.4 KB的UCRBP信息，与UCR结合因子的普遍表达相一致。将可表达的UCRBP cDNA瞬时转染到成纤维细胞中会导致MuLV启动子活性的下调，这与先前对UCR的功能分析一致。最近，三组已独立分离出人和小鼠UCRBP。这些研究表明，UCRBP结合了与UCR主题不同的各种目标基序：腺相关病毒P5启动子和免疫球蛋白轻链和重链基因中的元件，以及核糖体蛋白基因中的元件。这些结果表明UCRBP具有异常多样的DNA结合特异性，因此很可能调节许多不同基因的表达。

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