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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 顏伯勳(Bo-Shiun Yan) | |
dc.contributor.author | Tze-Chun Chen | en |
dc.contributor.author | 陳姿均 | zh_TW |
dc.date.accessioned | 2021-06-08T01:42:00Z | - |
dc.date.copyright | 2016-08-30 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-19 | |
dc.identifier.citation | 1. Muller, P.A. and K.H. Vousden, p53 mutations in cancer. Nat Cell Biol, 2013. 15(1): p. 2-8.
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Oncogene, 2002. 21(1): p. 9-21. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19000 | - |
dc.description.abstract | p53是個有很多重要細胞功能的蛋白,例如扮演腫瘤抑制子。面對各種壓力刺激時,p53會誘導特定的轉錄機轉,藉此調控細胞週期、細胞凋亡、細胞老化以及基因體的穩定。在細胞中,p53的表現和活性會被許多不同的後轉譯修飾,像是泛素化 (ubiquitination)、乙醯基化 (acetylation)、磷酸化 (phosphorylation)、甲基化 (methylation) 等所調控。不同類型的後轉譯修飾會參與p53的各類調控,有些會影響p53的穩定度,有些能夠調控活化基因表現的能力,有些則可能改變p53在細胞中的分佈。人類的SP110核質體蛋白可能參與調控基因轉錄、細胞死亡以及免疫反應,但目前對於其功能不是很清楚。實驗室先前的研究發現了SP110的其中一個異構物SP110b會與p53有交互作用。而且當SP110b大量表現時,p53在細胞內的分佈也會受到影響,使p53從原本分佈於整個細胞改變至只累積在細胞核內。進一步由液相層析串聯式質譜儀分析,發現SP110b的表現會使 p53上X 位點以及Y 位點出現被乙醯基化的現象,此現象可能會影響到p53的穩定度及活性。本論文於是進一步探討這兩個位點的乙醯基化對於p53可能的影響。實驗結果發現X或是Y位點的乙醯基化會增加p53的活性且改變p53在細胞內的分佈,尤其是Y位點乙醯基化所造成的影響更為明顯。然而,X或是Y位點的乙醯基化並不會影響到p53的穩定度。此外,p53 X及Y位點的乙醯基化在SP110b表現時對p53活性之影響會更加明顯。本論文的研究結果可以提供部份新的資訊解釋SP110b如何影響p53。 | zh_TW |
dc.description.abstract | p53, a protein with several cellular functions such as tumor suppressor, has well-established roles in monitoring various types of stress signals by activating specific transcriptional targets that control cell cycle, apoptosis, senescence as well as genomic DNA stability. In the cells, there are a multitude of post-translational modifications (PTMs), like ubiquitination, acetylation, methylation and phosphorylation, which could alter p53 protein level and activity in response to stresses. These different types of PTMs on p53 that usually influence protein stability, transcriptional activity or protein distribution may reflect different responses of p53. In human, SP110, a nuclear body protein, may involve in the regulation of transcriptional expression, cell death and immune response. However, the distinct functions should be further elucidated. In the previous work in the lab, we found that SP110b, one of SP110 major isoforms, has directly protein-protein interaction with p53. In the presence of SP110b overexpression, the localization of p53 was changed from the whole cell to the nucleus. Moreover, through LC MS/MS analysis, we found that SP110b expression induced acetylation of X and Y sites on p53 that may influence protein stability and activity. In this thesis, the studies were designed to determine whether the acetylation on X and Y sites could affect the functions of p53. The results demonstrated that the acetylation on the two sites affects p53 activity and cellular distribution and that, especially, the acetylation of Y site has stronger effects than those of X site. However, p53 stability doesn’t be affected by acetylation of X or Y site on p53. Besides, SP110b expression increased the X or Y site acetylation-mediated effect on p53 activity. The results of these studies provided a piece of new information to explain the relationship between SP110b and p53. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T01:42:00Z (GMT). No. of bitstreams: 1 ntu-105-R02442023-1.pdf: 3276077 bytes, checksum: 4c85b9e07c199ef0e1fa1e3c3403b36f (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 口試委員審定書……………………………………………………………..……#
致謝…………………………………………………………………………..……I 摘要……………………………………………………………………………….II Abstract………………………….…………………………………………….…III 目錄……………………………….…………………………………………...…IV 圖目錄………………………………………………………………………....…VI 表目錄……………………………………………………………………..….…VII 第一章緒論…………………………………………………………………….….1 1.1 p53………………...……………………………………………….……..….…1 1.2 SP110b………..………..…………………………………………….……...….3 1.3 研究目的……………………….……..………......………………..….………4 第二章材料與方法………………………………………………………….….….6 2.1質體 (Plasmids)………………………….……………...……………….…….6 2.2細胞株培養 (Cell culture)……………….……………...………………..……6 2.3 質體轉染 (Transfection)..………..………………….……...…………….…...7 2.4藥物處理 (Drug treatment)…...………..…………………..……………...…...7 2.5細胞蛋白之收取 (Cell lysis)…………...…….……………………..…….……8 2.6蛋白質之定量 (BCA protein assay)………..…………………………….…….9 2.7正十二烷硫酸鈉-聚丙烯醯胺膠體電泳 (SDS-PAGE)……..………….…...…9 2.8西方墨點法 (Western blot)…...………………………...……………….….....10 2.9冷光酵素報導基因分析 (Luciferase reporter assay)………..............…....….11 2.10免疫螢光分析 (Immunofluorescence Assay, IFA)…….………………….….11 2.11酵母菌雙雜交實驗 (Yeast two-hybrid)…….......…………………...…......…12 第三章實驗結果……………………………………..………………………… .....14 3.1以點突變模擬乙醯基化並進行質體的建構.……..……………..…….…. ..…14 3.2 p53 X及Y位點的乙醯基化對其活性造成的影響…………………..……….15 3.3 p53 X及Y位點的乙醯基化對其蛋白穩定度的影響………………….……..17 3.4 p53 X及Y位點的乙醯基化對其在細胞內分佈的影響……………...…....…18 3.5 p53 X及Y位點的乙醯基化對p53與SP110b之間交互作用的影響.............20 第四章討論…………………………………..……………..……………...….…….22 參考文獻…………...…………………………………………..…….…...……...….62 | |
dc.language.iso | zh-TW | |
dc.title | 乙醯基化及去乙醯基化對p53蛋白的調控 | zh_TW |
dc.title | The regulation of p53 by acetylation and deacetylation | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林敬哲,歐展言 | |
dc.subject.keyword | p53,SP110b,乙醯基化,後轉譯修飾,蛋白穩定度, | zh_TW |
dc.subject.keyword | p53,SP110b,Acetylation,Protein stability,Post-translational modification, | en |
dc.relation.page | 66 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2016-08-19 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
顯示於系所單位: | 生物化學暨分子生物學科研究所 |
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