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標題: | NF-κB 對大鼠懷孕特殊醣蛋白rnCGM3的基因調節所扮演的角色 Role of NF-κB in the expression of a rat pregnancy-specific glycoprotein gene, rnCGM3 |
作者: | Chia-Lin Liu 劉佳霖 |
出版年 : | 1998 |
學位: | 碩士 |
摘要: | 在哺乳動物的懷孕過程中,懷孕特殊醣蛋白(pregnancy-specific glycoprotein, PSG)是胎盤所表現的主要蛋白質。為了探討這些蛋白質的表達機制與生理功能,許多科學家找出了老鼠的相似基因並加以研究。其中一個基因,rnCGM3,其蛋白質序列由互補去氧核糖核酸序列推測,包含了三個相似於免疫球蛋白變異域(variable domain)及一個固定域(Constant domain)。由DNase I footprinting分析rnCGM3基因的啟動子與上游區域,鑑定出了三個調節區域(FPI, FPII, FPIII)。其中FPII區域已知受到NF-IL6的正向調節。繼上述的研究成果,我們進一步探討和FPIII區域作用的轉錄因數。藉由yeast one-hybrid system,我們發現其中一個是NF-КB。NF-КB已知在許多種細胞中都有表現,主要參與和免疫有關的反應中。在NF-КB的家族成員中,p50和FPIII的結合力最強,C-Rel與p65和FPIII的結合較弱。p65和FPIII的結合會促進基因表現,c-Rel相對較弱,p50則不會啟動FPIII調節的基因。C-Rel會協同p65強化FPIII報導基因的表現,而p50卻會減弱p65的活性。p65和FPII的結合因數NF-IL6之間有交互作用。我們的實驗結果顯示NF-КB協同NF-IL6共同調節rnCGM3的表現。 Pregnancy-specific glycoproteins (PSGs) are the most abundant proteins expressed by the placenta during pregnancy. To understand the physiological functions of PSG and the regulation of PSG gene expression during pregnancy, several rat homologues have been cloned and analyzed. One of them, rnCGM3, has three immunoglobulin (Ig) variable-like domain, and a Ig constant-like domain deduced from the cDNA sequence. The upstream region of rnCGM3 gene contains three regulatory elements (FPI, FPII, and FPIII) according to DNase I footprinting assay. The FPII region is positively regulated by NF-IL6. In this study, we further characterized the FPIII-binding factors using the yeast one- hybrid system. NF-КB, an ubiquitously expressed transcription factor which participates in the immune and inflammatory responses, was found to be one of the factors bound to FPIII. Electrophoretic mobility shift assay showed that FPIII has a higher binding affinity for p50 than c-Rel and p65. In a transient expression assay, we showed that both p65 and c-Rel could individually activate the FPIII-CAT reporter gene. A synergistic activation was observed when both p65 and c-Rel were present. p50 could not activate the FPIII-CAT reporter gene alone, however, it might compete for the FPIII site and reduce the p65 activation. We also demonstrated that an interaction between NF-IL6 and NE-КB is involved in the rnCGM3 expression. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76333 |
全文授權: | 未授權 |
顯示於系所單位: | 生化科學研究所 |
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