https://scholars.lib.ntu.edu.tw/handle/123456789/160414
標題: | 分析和c-Maf有交互作用之蛋白質研究以及c-Maf對IL-10之調控 Dissect c-Maf-interacting proteins and regulation of IL-10 by c-Maf |
作者: | 劉夢薇 Liu, Meng-Wei |
關鍵字: | c-Maf轉錄因子;c-Maf | 公開日期: | 2008 | 摘要: | 轉錄因子c-Maf是調控IL-4基因之特異性轉錄因子,並且在第二型輔助型T細胞(T helper 2 cells)的分化中扮演一個極為重要的角色。在被刺激之後,c-Maf的表現量會在Th2細胞以及巨噬細胞中大大提升,並引發大量的IL-4以及IL-10之表現,抑制IL-12的表達。因此,我們假設c-Maf之所以在不同的細胞中會有不同功能,可能是透過和不同的蛋白質有交互作用所致。用酵母菌雙雜交系統,我們實驗室已經篩選出SUMOylation的E2 conjugating enzyme Ubc9,E3 ligase PIAS1,以及蛋白酪氨酸磷酸脢PTPN22可能和c-Maf有交互作用。於是藉由螢光共振能量轉移的方法,再次確認了c-Maf的確跟這三個蛋白質有交互作用。另外一項假設是既然c-Maf可以和SUMOylation以及去磷酸脢等酵素有交互作用,暗示了c-Maf可以被SUMOylation以及磷酸化,而這些轉錄後修飾也許會對於c-Maf的轉譯能力造成影響。因此,藉由螢光酵素實驗,發現c-Maf的SUMOylation以及磷酸化突變種對於IL-10基因啟動子有較差的轉錄能力。 C-Maf is an IL-4-specific transcription factor that plays a vital role in Th2 cells differentiation. Upon stimulation, c-Maf is highly upregulated in Th2 cells as well as macrophages, and induces significant IL-4 and IL-10 expression but downregulates IL-12 expression. Thus, we hypothesized that c-Maf may have different functions through interacting with cell-specific proteins in different types of cells. Using yeast two-hybrid system, our group demonstrated that c-Maf can interact with Ubc9, the SUMOylation-specific E2 conjugating enzyme, PIAS1, the SUMOylation E3 ligase, and protein tyrosine phosphatase non-receptor type22 (PTPN22). Therefore, by Fluorescence Resonance Energy Transfer (FRET) assay, the interaction between c-Maf, Ubc9, c-Maf, PIAS1, and c-Maf, PTPN22 are confirmed. Since c-Maf can interact with SUMOylation enzymes and de-phosphorylation enzyme, which suggests that c-Maf undergoes SUMOylation and phosphorylation, these post-translational modification may affect the transactivity of c-Maf. Thereafter, we found that both c-Maf SUMOylation and phosphorylation mutants have poorer transactivity on IL-10 promoter by luciferase assay. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/181790 |
顯示於: | 免疫學研究所 |
檔案 | 描述 | 大小 | 格式 | |
---|---|---|---|---|
ntu-97-R95449005-1.pdf | 23.32 kB | Adobe PDF | 檢視/開啟 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。