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首页> 外文期刊>Molecular and Cellular Biology >Identification of an octamer-binding site in the mouse kappa light-chain immunoglobulin enhancer.
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Identification of an octamer-binding site in the mouse kappa light-chain immunoglobulin enhancer.

机译：小鼠κ轻链免疫球蛋白增强子中八聚体结合位点的鉴定。

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A 215-base-pair (bp) region of the mouse MOPC 41 kappa light-chain immunoglobulin gene enhancer has been analyzed for specific binding of lymphoid and nonlymphoid nuclear factors. Mobility shift assays with a series of overlapping DNA fragments have mapped DNA-binding sites for three unique factors. The B-cell-specific (OTF-2) and ubiquitous (OTF-1) octamer-binding transcription factors specifically bound to a site centered about 136 bp 5' of the nuclear factor NF-kappa B site. A third specific factor, NF-kappa E, bound to a site that was about 75 bp 5' of the NF-kappa B site and within a region important for enhancer function. This novel factor was found in both mature B and HeLa cell nuclei. B-cell OTF-2, B-cell OTF-1, and HeLa OTF-1 bound to the kappa enhancer and kappa promoter octamer sites with similar affinities despite a 2-bp difference in the kappa enhancer octamer sequence. However, DNase I footprint analyses indicated that affinity-purified OTF-2 bound both to the enhancer OTF site and, surprisingly, to 80 bp of A + T-rich flanking sequence. Moreover, methylation interference studies demonstrated distinct differences in OTF interactions between the consensus octamer in the kappa promoter and the nonconsensus octamer identified in the enhancer. This novel observation of an OTF-binding site in the kappa enhancer provides a common link with the OTF sites in the promoter-proximal regions of all kappa promoters and thus mirrors the structural arrangement of OTF sites found in the promoters and enhancers of immunoglobulin heavy-chain genes.
机译：小鼠MOPC 41 kappa轻链免疫球蛋白基因增强子的215个碱基对（bp）区域已被分析的淋巴和非淋巴核因子的特异性结合。具有一系列重叠DNA片段的迁移率迁移分析已绘制了三个独特因子的DNA结合位点。 B细胞特异性（OTF-2）和泛在（OTF-1）八聚体结合转录因子特异性结合至以核因子NF-κB位点约136 bp 5'为中心的位点。第三特定因子，NF-κB，结合到NF-κB位点的约75bp 5'的位点，并且在对于增强子功能重要的区域内。在成熟的B和HeLa细胞核中均发现了这一新因子。 B细胞OTF-2，B细胞OTF-1和HeLa OTF-1以相似的亲和力与κ增强子和κ启动子八聚体位点结合，尽管κ增强子八聚体序列有2 bp的差异。但是，DNase I足迹分析表明，亲和纯化的OTF-2既与增强子OTF位点结合，又与富A + T的侧翼序列的80 bp结合。此外，甲基化干扰研究表明，κ启动子中的共有八聚体与增强子中确定的非共识八聚体之间的OTF相互作用存在明显差异。对Kappa增强子中OTF结合位点的这种新颖观察提供了与所有Kappa启动子的启动子近端区域中的OTF位点的公共链接，因此反映了在免疫球蛋白重链启动子和增强子中发现的OTF位点的结构排列。链基因。

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