克隆人类基因的细胞间粘附分子1和5 '监管区域的分析。诱导细胞因子和佛波醇酯。
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Voraberger G,谢弗R, Stratowa C
克隆人类基因的细胞间粘附分子1和5 '监管区域的分析。诱导细胞因子和佛波醇酯。
J Immunol。1991年10月15日,147 (8):2777 - 86。
- PubMed ID
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1680919 (在PubMed]
- 文摘
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人类细胞间粘附molecule-1 (ICAM-1),淋巴细胞的特定配体关联Ag-1 (LFA-1)中扮演一个重要的角色在leukocyte-endothelial细胞相互作用。诱导的促炎细胞因子il - 1、tnf、IFN-gamma。然而,所知甚少涉及的细胞内调节机制触发ICAM-1老年病。为了研究潜在的监管元素参与ICAM-1感应我们克隆出人类ICAM-1基因的5 '监管区域和5 kb。互补脱氧核糖核酸被发现的序列分布在7个外显子相隔6个内含子,即五细胞外Ig-like ICAM-1域是由自己的外显子编码。上游序列港口大量的序列图案涉及监管和真核基因的表达,包括转录因子结合位点sp 1, AP-1, NF-kB。引物延伸和S1核酸酶分析发现两个转录起始位点319个基点,41岁的英国石油公司上游的翻译网站开始。共识TATA盒被发现在预期的位置大约25个基点的上游网站开始。逆转录酶聚合酶链反应显示,A549微分使用两个TATA框和HS913T细胞。两个RNA似乎相同的代码ICAM-1蛋白质。 For regulation studies a 1.3-kb EcoRI/SalI fragment of the 5'-flanking region was used to promote transcription of a linked luciferase reporter gene in transient-transfection assays in A549 and HS913T cells. Treatment of A549 cells with IL-1 or TNF-alpha resulted in a two- or fourfold increase in luciferase activity. Furthermore, a sixfold induction could be achieved after treatment with the phorbol ester PMA. In contrast, agents that increase intracellular cAMP levels did not induce luciferase activity. Northern blot analysis was used to investigate the kinetics of ICAM-1 mRNA synthesis upon induction with TNF-alpha and PMA. These data suggest that the up-regulation of ICAM-1 by cytokines occurs at least partly at the transcriptional level. Deletion analysis of the 1.3-kb fragment of the 5'-flanking region revealed sequences responsible for promotion and inhibition of transcription. In particular, two functionally distinct regions have been characterized: a short fragment containing an NF-kB binding site has been shown to function as an activator, followed immediately downstream by a sequence acting as a silencer element. Therefore, ICAM-1 gene expression seems to be modulated by multiple cis-acting elements.