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研究生(外文):William Tseng
論文名稱(外文):Regulation of IL-4 and IL-21 gene expression by the basic region of c-Maf
外文關鍵詞:c-Mafbasic regionIL-4IL-21
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在CD4輔助T細胞中,轉錄因子 c-Maf 是一重要的調控因子。c-Maf 會週控介白素4 (IL-4) 在第二型輔助T細胞 (TH2 cells) 和介白素21 (IL-21) 在17型輔助T細胞 (TH17 cells) 以及濾泡輔助型T細胞 (TFH cells) 中的基因表現。介白素4是第二型輔助T細胞的特徵細胞激素,並對於第二型輔助T細胞之功能有重要的影響。而 c-Maf 則藉由對介白素4的影響對第二型輔助T細胞的分化有重要的功能。最近,c-Maf 被發現也會表現在新發現的17型輔助T細胞以及濾泡輔助型T細胞中並調控介白素21的表現。介白素21對發炎反應和B細胞的生發中心反應 (Germinal center reaction) 有重要的影響。為了驗證 c-Maf 鹼性區域 (basic region) 在功能上的角色,我們利用了幾個在鹼性區域中的突變。我們發現鹼性區域在 c-Maf 調控介白素4上有重要性但在調控介白素21上則沒有影響。這些突變影響了 c-Maf 結合 DNA 的能力,但卻增強了 c-Maf 蛋白質的穩定度。而造成在 IL-4 和 IL-21 上有不同影響的機制則需要進一步的研究。

論文口試委員審定書 i
Acknowledgment ii
摘要 iii
Abstract iv
Chapter Ⅰ Introduction 1
1. CD4+ Helper T Cells 1
2. The Maf Family 3
3. Overview of Transcription Factor c-Maf 4
4. Transcription Factor c-Maf And Its Function In Immune System 6
5. Overviews of Cytokine IL-4 and IL-21 7
6. Overview of c-Maf Basic Region and Mutations in Basic Region 8
Chapter Ⅱ Materials and Methods 10
1. Experimental Materials 10
1.1 Chemicals and Reagents 10
1.2 Enzymes And Antibodies 12
1.3 Kits 12
1.4 Instruments 13
1.5 Medium, Solutions and Buffers 13
2. Experimental Procedure 15
2.1 Plasmid construction and preparation 15
2.2 Cell Culture and Transfection 16
2.3 Luciferase Assay 17
2.4 Western Blotting Analysis 17
2.5 Nucleus and Cytoplasmic Fractionation 17
2.6 Electrophoretic Mobility Shift Assay (EMSA) 17
2.7 Confocal Micorscopy 18
2.8 Stability Assay 19
Chapter Ⅲ Results 20
1. c-Maf with mutation in basic region affects its transactivity on IL-4 promoter but not IL-21 promoter. 20
2. The DNA binding abilities of Mutation A, Mutation B and Mutation C are impaired. 21
3. The cellular localization of Mutation A, Mutation B and Mutation C c-Maf remains in the nucleus. 22
4. c-Maf Mutation A, Mutation B and Mutation C is more stable than wild-type. 23
5. The DNA binding domain of c-Maf is indispensable in IL-21 transactivation. 23
6. Batf does not cooperate with c-Maf to regulate IL-21 expression. 24
Chapter Ⅳ Discussion 26
1. The c-Maf basic region plays significant roles in DNA binding and MARE recognition of c-Maf Proteins. 26
2. c-Maf may cooperate with other protein to regulate IL-21 expression. 27
3. The c-Maf instability may be related to DNA binding and regulation of gene expression control. 29
4. The subcellular localization of Mutation A, Mutation B and Mutation C c-Maf need to be further examined. 29
Figures 31
Reference 45

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