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Overlapping Expression of Early B-Cell Factor and Basic Helix-Loop-Helix Proteins as a Mechanism To Dictate B-Lineage-Specific Activity of the λ5 Promoter

机译:重叠的早期B细胞因子和基本的螺旋环螺旋蛋白的表达作为决定λ5启动子B谱系特异性活性的机制

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The basic helix-loop-helix (bHLH) transcription factors are a large group of proteins suggested to control key events in the development of B lymphocytes as well as of other cellular lineages. To examine how bHLH proteins activate a B-lineage-specific promoter, I investigated the ability of E47, E12, Heb, E2-2, and MyoD to activate the λ5 surrogate light chain promoter. Comparison of the functional capacity of the E2A-encoded E47 and E12 proteins indicated that even though both were able to activate the λ5 promoter and act in synergy with early B-cell factor (EBF), E47 displayed a higher functional activity than E12. An ability to act in synergy with EBF was also observed for Heb, E2-2, and MyoD, suggesting that these factors were functionally redundant in this regard. Mapping of functional domains in EBF and E47 revealed that the dimerization and DNA binding domains mediated the synergistic activity. Electrophoretic mobility shift assay analysis using the 5′ part of the λ5 promoter revealed formation of template-dependent heteromeric complexes between EBF and E47, suggesting that the synergistic mechanism involves cooperative binding to DNA. These findings propose a unique molecular function for E47 and provide overlapping expression with EBF as a molecular mechanism to direct B-cell-specific target gene activation by bHLH proteins.
机译:基本的螺旋-环-螺旋(bHLH)转录因子是一大类蛋白质,被认为可控制B淋巴细胞以及其他细胞谱系发育中的关键事件。为了检查bHLH蛋白如何激活B谱系特异性启动子,我研究了E47,E12,Heb,E2-2和MyoD激活λ 5 替代轻链启动子的能力。对 E2A 编码的E47和E12蛋白的功能能力进行比较表明,即使两者都能够激活λ 5 启动子并与早期B细胞协同作用因子(EBF),E47的功能活性高于E12。还观察到了Heb,E2-2和MyoD与EBF协同作用的能力,这表明这些方面在功能上是多余的。 EBF和E47中的功能域映射显示,二聚化和DNA结合域介导了协同活性。使用λ 5 启动子的5'部分进行的电泳迁移率变动分析表明,EBF和E47之间存在模板依赖性异源复合物,这表明协同机制涉及与DNA的协同结合。这些发现为E47提出了独特的分子功能,并提供了与EBF的重叠表达,作为指导bHLH蛋白激活B细胞特异性靶基因的分子机制。

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