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The regulation of CD5 expression in murine T cells

机译:小鼠T细胞CD5表达的调控。

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Background CD5 is a pan-T cell surface marker that is also present on a subset of B cells, B-1a cells.Functional and developmental subsets of T cells express characteristic CD5 levels that vary over roughly a 30-fold range. Previous investigators have cloned a 1.7 Kb fragment containing the CD5 promoter and showed that it can confer similar lymphocyte-specific expression pattern as observed for endogenous CD5 expression. Results We further characterize the CD5 promoter and identify minimal and regulatory regions on the CD5 promoter. Using a luciferase reporter system, we show that a 43 bp region on the CD5 promoter regulates CD5 expression in resting mouse thymoma EL4 T cells and that an Ets binding site within the 43 bp region mediates the CD5 expression. In addition, we show that Ets-1, a member of the Ets family of transcription factors, recognizes the Ets binding site in the electrophoretic mobility shift assay (EMSA). This Ets binding site is directly responsible for the increase in reporter activity when co-transfected with increasing amounts of Ets-1 expression plasmid. We also identify two additional evolutionarily-conserved regions in the CD5 promoter (CD5X and CD5Y) and demonstrate the respective roles of the each region in the regulation of CD5 transcription. Conclusion Our studies define a minimal and regulatory promoter for CD5 and show that the CD5 expression level in T cells is at least partially dependent on the level of Ets-1 protein. Based on the findings in this report, we propose a model of CD5 transcriptional regulation in T cells.
机译:背景CD5是泛T细胞表面标志物,也存在于B细胞的一个子集B-1a细胞中.T细胞的功能和发育子集表达的特征性CD5水平大约在30倍范围内变化。先前的研究人员已经克隆了一个包含CD5启动子的1.7 Kb片段,并表明它可以赋予与内源性CD5表达相似的淋巴细胞特异性表达模式。结果我们进一步表征CD5启动子,并确定CD5启动子上的最小和调控区域。使用萤光素酶报告系统,我们显示CD5启动子上的43 bp区域调节了静止的小鼠胸腺瘤EL4 T细胞中CD5的表达,并且43 bp区域内的Ets结合位点介导了CD5的表达。此外,我们显示Ets-1,Ets家族的转录因子的成员,在电泳迁移率变动分析(EMSA)中识别Ets结合位点。当与增加量的Ets-1表达质粒共转染时,此Ets结合位点直接负责报告基因活性的增加。我们还确定了CD5启动子(CD5X和CD5Y)中两个其他的进化保守区域,并证明了每个区域在CD5转录调控中的各自作用。结论我们的研究确定了CD5的最小启动子和调节启动子,并表明T细胞中CD5的表达水平至少部分取决于Ets-1蛋白的水平。基于本报告中的发现,我们提出了T细胞中CD5转录调控的模型。

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