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首页> 外文期刊>Molecular and Cellular Biology >c-Myb and core-binding factor/PEBP2 display functional synergy but bind independently to adjacent sites in the T-cell receptor delta enhancer.
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c-Myb and core-binding factor/PEBP2 display functional synergy but bind independently to adjacent sites in the T-cell receptor delta enhancer.

机译：c-Myb和核心结合因子/ PEBP2显示功能协同作用，但独立地与T细胞受体δ增强子中的相邻位点结合。

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A T-cell-specific transcriptional enhancer lies within the J delta 3-C delta intron of the human T-cell receptor delta gene. We have previously shown that a 30-bp element, denoted delta E3, acts as the minimal TCR delta enhancer and that within delta E3, adjacent and precisely spaced binding sites for core-binding factor (CBF/PEBP2) and c-Myb are essential for transcriptional activity. These data suggested that CBF/PEBP2 and c-Myb synergize to mediate transcriptional activity but did not establish the molecular basis for synergy. In this study, we have examined in detail the binding of CBF/PEBP2 and c-Myb to delta E3. We found that CBF/PEBP2 and c-Myb could simultaneously occupy the core site and one of two overlapping Myb sites within delta E3. However, equilibrium binding and kinetic dissociation experiments suggest that the two factors bind to delta E3 independently, rather than cooperatively. This was found to be true by using isoforms of these factors present in extracts of transfected COS-7 cells, as well as the natural factors present in nuclear extracts of the Jurkat T-cell line. We further showed that CBF/PEBP2 and c-Myb provide unique transactivation functions, since the core-Myb combination cannot be substituted by dimerized core or Myb sites. We propose that spatially precise synergy between CBF/PEBP2 and c-Myb may result from the ability of the two factors to form a composite surface that makes unique and stereospecific contacts with one or more additional components of the transcriptional machinery.
机译：T细胞特异性转录增强子位于人T细胞受体delta基因的J delta 3-C delta内含子内。先前我们已经表明，一个30 bp的元件（称为delta E3）充当最小的TCR delta增强子，而在delta E3中，核心结合因子（CBF / PEBP2）和c-Myb的相邻且精确间隔的结合位点是必不可少的转录活性。这些数据表明CBF / PEBP2和c-Myb协同介导转录活性，但没有建立协同作用的分子基础。在这项研究中，我们详细检查了CBF / PEBP2和c-Myb与δE3的结合。我们发现CBF / PEBP2和c-Myb可以同时占据核心位点和δE3内两个重叠的Myb位点之一。然而，平衡结合和动力学解离实验表明这两个因素独立地而不是协同地结合到δE3上。通过使用转染的COS-7细胞提取物中存在的这些因子的同工型以及Jurkat T细胞系核提取物中存在的天然因子，发现这是正确的。我们进一步表明CBF / PEBP2和c-Myb提供了独特的反式激活功能，因为core-Myb组合不能被二聚化的core或Myb位点取代。我们提出，CBF / PEBP2和c-Myb之间的空间精确协同作用可能是由于这两个因素形成复合表面的能力所致，该复合表面与转录机制的一个或多个其他成分形成独特且立体的接触。

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