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首页> 外文期刊>Molecular and Cellular Biology >A central role for a single c-Myb binding site in a thymic locus control region.
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A central role for a single c-Myb binding site in a thymic locus control region.

机译：胸腺基因座控制区中单个c-Myb结合位点的核心作用。

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Locus control regions (LCRs) are powerful assemblies of cis elements that organize the actions of cell-type-specific trans-acting factors. A 2.3-kb LCR in the human adenosine deaminase (ADA) gene first intron, which controls expression in thymocytes, is composed of a 200-bp enhancer domain and extended flanking sequences that facilitate activation from within chromatin. Prior analyses have demonstrated that the enhancer contains a 28-bp core region and local adjacent augmentative cis elements. We now show that the core contains a single critical c-Myb binding site. In both transiently cotransfected human cells and stable chromatin-integrated yeast cells, c-Myb strongly transactivated reporter constructs that contained polymerized core sequences. c-Myb protein was strongly evident in T lymphoblasts in which the enhancer was active and was localized within discrete nuclear structures. Fetal murine thymus exhibited a striking concordance of endogenous c-myb expression with that of mouse ADA and human ADA LCR-directed transgene expression. Point mutation of the c-Myb site within the intact 2.3-kb LCR severely attenuated enhancer activity in transfections and LCR activity in transgenic thymocytes. Within the context of a complex enhancer and LCR, c-Myb can act as an organizer of thymocyte-specific gene expression via a single binding site.
机译：基因座控制区（LCR）是顺式元件的强大组装，可组织细胞类型特异性反式作用因子的作用。在人腺苷脱氨酶（ADA）基因第一内含子中控制着胸腺细胞表达的2.3 kb LCR由200 bp的增强子域和扩展的侧翼序列组成，这些序列有助于从染色质内部激活。先前的分析表明，该增强子包含一个28 bp的核心区域和局部相邻的增强型顺式元件。现在我们显示该核心包含一个关键的c-Myb结合位点。在瞬时共转染的人类细胞和稳定的染色质整合的酵母细胞中，c-Myb都强烈转导了含有聚合核心序列的报告基因构建体。 c-Myb蛋白在增强活性的T淋巴细胞中很明显，并且位于离散的核结构中。胎鼠胸腺表现出内源性c-myb表达与小鼠ADA和人ADA LCR定向转基因表达的惊人一致性。完整的2.3-kb LCR中c-Myb位点的点突变严重减弱了转染中的增强子活性和转基因胸腺细胞中的LCR活性。在复杂的增强子和LCR的背景下，c-Myb可以通过单个结合位点充当胸腺细胞特异性基因表达的组织者。

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