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首页> 外文期刊>Molecular and Cellular Biology >Binding of multiple nuclear factors to the 5' upstream regulatory element of the murine major histocompatibility class I gene.
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Binding of multiple nuclear factors to the 5' upstream regulatory element of the murine major histocompatibility class I gene.

机译：多种核因子与鼠类主要组织相容性I类基因的5'上游调控元件的结合。

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Transcription of mouse major histocompatibility complex class I genes is controlled by the conserved class I regulatory element (CRE) in the 5' flanking region. The CRE, approximately 40 base pairs long, acts as a negative control element in undifferentiated F9 embryonal carcinoma cells which do not express the major histocompatibility complex genes. The same element, however, acts as a positive control element in cells expressing the genes at high levels. To investigate the molecular basis of the regulatory role of the CRE, we studied the binding of nuclear proteins to the CRE of the H-2Ld gene by gel mobility shift and methylation interference experiments. Nuclear extracts from L fibroblasts and LH8 T lymphocytes revealed three distinct factors that bind discrete sequences within the CRE. The three sequences correspond to the inverted and direct repeats within the CRE. In contrast, F9 extracts exhibited factor binding to only two of the three sequences and lack a major factor detected in the above two cell types. Protein-binding sites within each of the three sequences were identified by methylation interference experiments. These data were in full agreement with results obtained by a competition assay performed with a series of mutant oligonucleotides containing a few nucleotide substitutions in each of the three regions. The results illustrate complex DNA-protein interactions in which several independent proteins bind to overlapping sequences in the CRE in a cell type-specific fashion.
机译：小鼠主要组织相容性复杂的I类基因的转录受5'侧翼区域保守的I类调节元件（CRE）的控制。 CRE长约40个碱基对，在未分化的F9胚胎癌细胞中充当阴性控制元件，这些细胞不表达主要的组织相容性复合物基因。然而，相同的元件在高水平表达基因的细胞中充当阳性对照元件。为了研究CRE调控作用的分子基础，我们通过凝胶迁移率迁移和甲基化干扰实验研究了核蛋白与H-2Ld基因CRE的结合。 L成纤维细胞和LH8 T淋巴细胞的核提取物揭示了三个不同的因素，它们结合了CRE中的离散序列。这三个序列对应于CRE内的反向和直接重复。相反，F9提取物仅与三个序列中的两个序列显示因子结合，而在上述两种细胞类型中缺乏检测到的主要因子。通过甲基化干扰实验鉴定了三个序列中每个序列内的蛋白质结合位点。这些数据与通过竞争分析获得的结果完全一致，该竞争分析是用一系列突变寡核苷酸进行的，该突变寡核苷酸在三个区域的每一个中都包含一些核苷酸取代。结果说明了复杂的DNA-蛋白质相互作用，其中几种独立的蛋白质以细胞类型特异性方式与CRE中的重叠序列结合。

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