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小鼠T细胞中CD5表达的调控。

The regulation of CD5 expression in murine T cells.

作者信息

Tung J W, Kunnavatana S S, Herzenberg L A, Herzenberg L A

机构信息

Department of Genetics, Stanford University Medical School, Stanford, CA 94305, USA.

出版信息

BMC Mol Biol. 2001;2:5. doi: 10.1186/1471-2199-2-5. Epub 2001 May 22.

Abstract

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.

摘要

背景

CD5是一种泛T细胞表面标志物,也存在于一部分B细胞即B-1a细胞上。T细胞的功能和发育亚群表达特征性的CD5水平,其变化范围大致为30倍。先前的研究者克隆了一个包含CD5启动子的1.7 Kb片段,并表明它可以赋予与内源性CD5表达所观察到的相似的淋巴细胞特异性表达模式。

结果

我们进一步对CD5启动子进行了表征,并确定了CD5启动子上的最小区域和调控区域。使用荧光素酶报告系统,我们表明CD5启动子上一个43 bp的区域调节静息小鼠胸腺瘤EL4 T细胞中的CD5表达,并且该43 bp区域内的一个Ets结合位点介导CD5表达。此外,我们表明转录因子Ets家族的成员Ets-1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,这个Ets结合位点直接导致报告基因活性增加。我们还在CD5启动子中鉴定出另外两个进化保守区域(CD5X和CD5Y),并证明了每个区域在CD5转录调控中的各自作用。

结论

我们的研究定义了CD5的最小调控启动子,并表明T细胞中CD5的表达水平至少部分依赖于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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bce8/32207/b6790f655e08/1471-2199-2-5-1.jpg

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