Viville S, Jongeneel V, Koch W, Mantovani R, Benoist C, Mathis D
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génetique de l'INSERM, Faculté de Médecine, Strasbourg, France.
J Immunol. 1991 May 1;146(9):3211-7.
To accurately delineate the DNA sequence elements involved in the transcriptional regulation of the murine MHC class II gene E alpha, we have constructed a set of "linker-scanning" mutants of the E alpha promoter region. These were made by the gapped heteroduplex technique and result in a set of 10-bp replacement mutations, covering the -206 to -6 stretch of the promoter. The effect of these mutations on transcriptional activity was evaluated in several systems, either by transfection into cultured cells or by in vitro transcription. The data points to the now classical X and Y boxes as the most important control elements, either for constitutive expression in B lymphoma cells, or for IFN-gamma-inducible expression in macrophages. Motifs upstream of the X box also play a role, but are somewhat less critical. Overall, we find no marked difference between regulatory strategies in B lymphomas or activated macrophages.
为了准确描绘参与小鼠MHC II类基因Eα转录调控的DNA序列元件，我们构建了一组Eα启动子区域的“接头扫描”突变体。这些突变体通过缺口异源双链技术构建，产生了一组10个碱基对的替换突变，覆盖启动子的-206至-6区域。通过转染到培养细胞中或体外转录，在多个系统中评估了这些突变对转录活性的影响。数据表明，现在经典的X盒和Y盒是最重要的控制元件，无论是对于B淋巴瘤细胞中的组成型表达，还是对于巨噬细胞中IFN-γ诱导型表达而言。X盒上游的基序也发挥作用，但重要性稍低。总体而言，我们发现在B淋巴瘤或活化巨噬细胞中的调控策略之间没有明显差异。
