Ghersa P, Hooft van Huijsduijnen R, Whelan J, DeLamarter J F
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
J Biol Chem. 1992 Sep 25;267(27):19226-32.
Endothelial leukocyte adhesion molecule-1 (ELAM-1) is a membrane protein exclusively expressed on endothelial cells, where it plays a key role in the inflammatory response by adhering to a subset of leukocytes. The expression of the ELAM-1 gene is very tightly regulated. ELAM-1 is undetectable in uninduced cells, and it is transiently expressed following cytokine induction. Treatment of resting endothelial cells with three different protein synthesis inhibitors, cycloheximide (CHX), anisomycin, and emetine, caused an increase in the steady-state level of ELAM-1 mRNA above that observed with IL (interleukin)-1 alone. Furthermore, ELAM-1 mRNA was found in the presence of all three protein synthesis inhibitors without IL-1 treatment. Analysis of the mRNA half-life indicated that the protein synthesis inhibitors act, in part, by stabilizing ELAM-1 mRNA. In addition, protein synthesis inhibitors potentiate the effect of IL-1 beta at the level of transcription initiation as shown by nuclear run-on experiments. The NF kappa B-like binding activity to the ELAM-1 promoter sequence induced by IL-1 beta is augmented by inhibitors of protein synthesis. The NF kappa B binding sequence was found to be necessary and sufficient for superinduction of the ELAM-1 gene by CHX. These results show that regulation at the level of protein synthesis is implicated in the overall regulation of ELAM-1 gene expression. Mechanisms which could explain these effects are discussed.
内皮细胞白细胞黏附分子-1(ELAM-1)是一种仅在内皮细胞上表达的膜蛋白,它通过黏附于一部分白细胞在炎症反应中发挥关键作用。ELAM-1基因的表达受到非常严格的调控。在未诱导的细胞中检测不到ELAM-1,在细胞因子诱导后它会短暂表达。用三种不同的蛋白质合成抑制剂环己酰亚胺(CHX)、茴香霉素和依米丁处理静息的内皮细胞,导致ELAM-1 mRNA的稳态水平高于单独用白细胞介素(IL)-1处理时观察到的水平。此外,在未用IL-1处理但存在所有三种蛋白质合成抑制剂的情况下也发现了ELAM-1 mRNA。对mRNA半衰期的分析表明,蛋白质合成抑制剂部分通过稳定ELAM-1 mRNA起作用。另外,如核转录实验所示,蛋白质合成抑制剂在转录起始水平增强了IL-1β的作用。蛋白质合成抑制剂增强了由IL-1β诱导的与ELAM-1启动子序列的NFκB样结合活性。发现NFκB结合序列对于CHX对ELAM-1基因的超诱导是必要且充分的。这些结果表明,蛋白质合成水平的调控参与了ELAM-1基因表达的整体调控。讨论了解释这些效应的机制。