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系統識別號 U0026-2907201414035300
論文名稱(中文) 鑑別與微小球蛋白(MSP58)相互結合之蛋白質
論文名稱(英文) Identification of a novel interacting partner of 58-kDa microspherule protein (MSP58)
校院名稱 成功大學
系所名稱(中) 生物資訊與訊息傳遞研究所
系所名稱(英) Insitute of Bioinformatics and Biosignal Transduction
學年度 102
學期 2
出版年 103
研究生(中文) 龔建融
研究生(英文) Chien-Jung Kung
學號 z26011039
學位類別 碩士
語文別 中文
論文頁數 67頁
口試委員 指導教授-林鼎晏
口試委員-張文昌
口試委員-陳昶翰
中文關鍵字 MSP58  RING指環  TRIM蛋白家族  小泛素化  泛素化 
英文關鍵字 MSP58  RING finger gene  TRIM family proteins 
學科別分類
中文摘要 58-kDa微小球蛋白(MSP58)最早發現和細胞增生相關的核仁蛋白p120能直接的蛋白質交互作用。早期針對MSP58的研究指出其功能在基因轉路調控與促進細胞轉型上扮演了重要角色。我們利用酵母菌雙雜交篩選技術找到了數個新穎的MSP58結合蛋白。 其中有一為具有RING指環,且屬於TRIM家族蛋白的成員。我們首先在哺乳類細胞透過co-immunoprecipitation(co-IP)的方式證實MSP58與此TRIM相關蛋白在體內(in vivo)會有蛋白質交互作用。TIRM家族蛋白屬於scaffold類蛋白並且參與在調控其他蛋白的類泛素化(sumoylation)及泛素化(ubiquitination)中。利用免疫螢光染色的結果發現此TRIM相關蛋白座落在細胞核且會將MSP58招集至核內特定區域(nuclear speckles)。進一步發現MSP58蛋白質可以調節此TRIM相關蛋白所調控的基因轉錄。值得注意的是,過去研究也指出其他TRIM蛋白家族成員,如TRIM27,能在核仁內與MSP58以及upstream binding factor(UBF) 形成複合體,且能直接參與調控核醣體基因的轉錄。未來的研究中,我們需要釐清MSP58與此TRIM相關蛋白複合體在蛋白質後轉譯修飾及基因轉錄調控的分子機制。
英文摘要 The 58-kDa microspherule protein (MSP58) was initially shown to interact with the nu-cleolar protein p120, a proliferation-associated protein. Earlier studies had proposed a role for MSP58 in the regulation of gene transcription as well as in cellular transfor-mation. We conducted yeast two-hybrid screening using MSP58 as a bait and identified several interacting partners. Here, we identified a novel RING finger gene, which en-codes a protein identical to TRIM family proteins as a MSP58-binding protein. The in-teraction between MSP58 and this proteinwas further validated by co-immunoprecipitation assays in mammalian cells. TRIM proteins could form the scaf-fold and define compartment to host many proteins involved in sumoylation or ubiquiti-nation. Immunofluorescence staining indicated that this protein recruits MSP58 to nu-clear speckles. We further demonstrated that MSP58 regulates this protein-mediated transcription attenuation. Notably, another TRIM family protein, TRIM27, have been reported to form a complex with MSP58 and UBF in the nucleolus, where they play a direct transactivating role on ribosomal gene transcription. Further studies are required to understand the molecular mechanisms of MSP58 and this protein complex in protein post-translational modifications and transcriptional regulation.
論文目次 中文摘要 I
英文摘要 II
致謝 V
目錄 VI
圖目錄 VII
附錄目錄 VIII
縮寫指引 IX
第一章 緒論 1
第二章 實驗方法與材料 13
第三章 實驗結果 39
第四章 討論 46
第五章 參考文獻 50
附圖 56
附錄 67
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