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首页> 外文期刊>Cytokine >Human interleukin-1alpha gene expression is regulated by Sp1 and a transcriptional repressor.
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Human interleukin-1alpha gene expression is regulated by Sp1 and a transcriptional repressor.

机译：人白介素-1α基因表达受Sp1和转录阻遏物调控。

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摘要

The regulation of the human IL-1alpha gene was studied using a series of 5' deletion promoter chloramphenicol acetyltransferase (CAT) reporter constructs. The IL-1alpha promoter from -967 to +64 produced no significant expression of CAT. Progressive 5' deletion indicated the presence of a repressor binding site between -477 and -305 bp as deletion in this region resulted in CAT expression. Electrophoretic mobility shift assay (EMSA) analysis confirmed that protein(s) bound to this region and DNaseI footprinting localized the binding site to between -448 and -435. Deletion of the IL-1alpha promoter to -42 resulted in reduced CAT expression suggesting the presence of a positive regulatory element in this region. EMSA experiments using IL-1alpha promoter DNA from -163 to +64 demonstrated protein binding to this region and DNaseI footprinting demonstrated protection between -59 and -40. Transcriptional activity of the IL-1alpha promoter was also tested using an in vitro transcription assay. Reactions using-163, -100 and -52 promoter templates all produced a correctly sized transcript but deletion to -42 resulted in no transcript production. Analysis of the promoter indicated that a potential Sp1 binding site existed in the region from -52 to -45. An EMSA using an anti-Sp1 antibody indicated that Sp1 specifically bound to the -52 to +64 region.
机译：使用一系列5'缺失启动子氯霉素乙酰转移酶（CAT）报告基因构建体研究了人类IL-1alpha基因的调控。从-967到+64的IL-1alpha启动子没有产生明显的CAT表达。渐进的5'缺失表明在-477和-305 bp之间存在阻遏物结合位点，因为该区域的缺失导致CAT表达。电泳迁移率变动分析（EMSA）分析证实，结合到该区域的蛋白质和DNaseI足迹将结合位点定位在-448和-435之间。 IL-1α启动子缺失至-42导致CAT表达降低，表明该区域存在正调控元件。使用从-163到+64的IL-1alpha启动子DNA进行的EMSA实验表明，蛋白质与该区域结合，而DNaseI足迹显示出了在-59和-40之间的保护。还使用体外转录测定法测试了IL-1α启动子的转录活性。使用-163，-100和-52启动子模板的反应均产生正确大小的转录本，但缺失至-42则不会产生任何转录本。启动子的分析表明，潜在的Sp1结合位点存在于-52至-45区域。使用抗Sp1抗体的EMSA表明Sp1特异性结合-52至+64区域。

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