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首页> 外文期刊>Biochemistry >Cyclic AMP response element-binding protein (CREB) and CAAT enhancer-binding protein, beta (C/EBP beta) bind chimeric DNA sites with high affinity
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Cyclic AMP response element-binding protein (CREB) and CAAT enhancer-binding protein, beta (C/EBP beta) bind chimeric DNA sites with high affinity

机译：环状AMP反应元件结合蛋白（CREB）和CAAT增强子结合蛋白beta（C / EBP beta）以高亲和力结合嵌合DNA位点

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摘要

Basic region leucine zipper (bZIP) proteins are transcription factors that interact selectively with duplex DNA to regulate gene expression. Specifically, the cAMP response element-binding protein (CREB) interacts with the cAMP response element (CRE) DNA site with high affinity, while it binds the CAAT/enhancer-binding protein (CEBP) DNA site with low affinity. Despite the selectivity of CREB for the CRE site, CREB-dependent transcription is observed via chimeric DNA sites with similarities to both CRE and CEBP sites. Because CRE/CEBP and CEBP/CRE chimeric DNA are relevant for transcription regulation but have not been rigorously characterized, quantitative electrophoretic mobility shift assays were used to characterize the binding affinity and specificity of CREB to the sites. In addition to CREB, C/EBP beta was tested because chimeric DNA was shown to stabilize CREB-C/EBP beta heterodimerization. Despite previous work, no CREB-C/EBP beta heterodimer was observed in the presence of chimeric DNA; only CREB and C/EBP beta homodimers were seen. The CREB homodimer bound to the chimeric sites with high affinity, demonstrating that the presence of one CRE half-site is sufficient for high-affinity interaction. A comparison of CREB and C/EBP, homodimers indicated that they bind the chimeric sites with similar, high affinity. Whereas the CRE and CEBP sites preferentially interact with CREB and C/EBP beta, respectively, the chimeric sites bind CREB and C/EBP beta competitively. Because DNA binding correlates with transcription regulation, the results suggest that gene expression from chimeric sites can be altered by small changes in relative bZIP concentrations or bZIP accessory factors.
机译：基本区域亮氨酸拉链（bZIP）蛋白是与双链DNA选择性相互作用以调节基因表达的转录因子。具体而言，cAMP反应元件结合蛋白（CREB）与cAMP反应元件（CRE）DNA位点相互作用，而亲和力高，而与CAAT /增强子结合蛋白（CEBP）DNA位点结合，亲和力低。尽管CREB对CRE位点具有选择性，但可通过与CRE和CEBP位点相似的嵌合DNA位点观察到CREB依赖性转录。因为CRE / CEBP和CEBP / CRE嵌合DNA与转录调控有关，但尚未得到严格的表征，所以使用定量电泳迁移率变动分析法来表征CREB与位点的结合亲和力和特异性。除CREB外，还测试了C / EBP beta，因为显示嵌合DNA可以稳定CREB-C / EBP beta异二聚体。尽管有以前的工作，但在嵌合DNA的存在下，未观察到CREB-C / EBPβ异二聚体。仅观察到CREB和C / EBPβ同型二聚体。 CREB同二聚体以高亲和力结合到嵌合位点，表明存在一个CRE半位足以实现高亲和力相互作用。 CREB和C / EBP同型二聚体的比较表明，它们以相似的高亲和力结合嵌合位点。 CRE和CEBP位点分别优先与CREB和C / EBP beta相互作用，而嵌合位点则竞争性地结合CREB和C / EBP beta。因为DNA结合与转录调控相关，所以结果表明嵌合位点的基因表达可以通过相对bZIP浓度或bZIP辅助因子的微小变化而改变。

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