Identification of two regulatory elements controlling Fucosyltransferase 7 transcription in murine CD4+ T cells.

Identification of two regulatory elements controlling Fucosyltransferase 7 transcription in murine CD4+ T cells.
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鼠 CD4 T 细胞中控制岩藻糖基转移酶 7 转录的两个调控元件的鉴定

DOI:
10.1016/j.molimm.2014.05.005
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发表时间:
2014
影响因子:
3.6
通讯作者:
U. Syrbe
U. Syrbe
中科院分区:
医学3区
文献类型:
--
作者:
B. A. Ratsch;M. Mardahl;M. F. Schroter;D. Engelbert;J. Triebus;A. Hamann;U. Syrbe

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由fut7基因编码的focusyltransferase VII在CD4+T细胞中产生E-和p -选择素配体(E-和p - light)是必不可少的,E-和p - light有助于淋巴细胞募集到炎症组织和皮肤中。本研究旨在鉴定CD4+T细胞活化并分化为效应细胞时可诱导fut7表达的调控元件。对非造血细胞和CD4+T细胞亚群的组蛋白修饰模式的比较分析显示,fut7位点存在差异的组蛋白修饰模式,包括一个保守的非编码序列(CNS),这表明调节元件仅限于该区域。将位于fut7位点上游约500 bp处的CNS克隆到荧光素酶报告载体中,转染αβ-WT T细胞系后可引起报告活性,但转染原代小鼠CD4+Th1细胞后无报告活性。由于对不同fut7转录本的定量分析显示,在原代Th1细胞中缺乏第一外显子的转录本占主导地位,我们寻找一个替代的启动子。将4号外显子上游1kb的基因内区域克隆为含有增强子的载体,确实可以激发启动子活性。有趣的是,在原代Th1细胞的报告基因试验中,CNS也增强了这种基因内最小启动子的活性,这表明这两种元件在原代CD4+T细胞中相互作用,诱导fut7转录。
Fucosyltransferase VII encoded by the geneFut7is essential in CD4+T cells for the generation of E- and P-selectin ligands (E- and P-lig) which facilitate recruitment of lymphocytes into inflamed tissues and into the skin.This study aimed to identify regulatory elements controlling the inducibleFut7expression in CD4+T cells that occurs upon activation and differentiation of naive T cells into effector cells.Comparative analysis of the histone modification pattern in non-hematopoetic cells and CD4+T cell subsets revealed a differential histone modification pattern within theFut7locus including a conserved non-coding sequence (CNS) identified by cross-species conservation comparison suggesting that regulatory elements are confined to this region.Cloning of the CNS located about 500 bp upstream of theFut7locus, into a luciferase reporter vector elicited reporter activity after transfection of the αβ-WT T cell line, but not after transfection of primary murine CD4+Th1 cells. As quantification of differentFut7transcripts revealed a predominance of transcripts lacking the first exons in primary Th1 cells we searched for an alternative promoter. Cloning of an intragenic region spanning a 1 kb region upstream of exon 4 into an enhancer-containing vector indeed elicited promoter activity. Interestingly, also the CNS enhanced activity of this intragenic minimal promoter in reporter assays in primary Th1 cells suggesting that both elements interact in primary CD4+T cells to induceFut7transcription.
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