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標題: | 一段潛藏的核仁定位訊號在SP110蛋白功能的作用 The role of a cryptic nucleolar-localization signal in SP110 protein function |
其他標題: | The role of a cryptic nucleolar-localization signal in SP110 protein function |
作者: | 顏廷涓 Ting-Chuan Yen |
指導教授: | 顏伯勳 Bo-Shiun Yan |
關鍵字: | SP110,細胞定位,核定位訊號,核仁定位訊號,微氧壓力, SP110,Cellular localization,Nuclear localization signal (NLS),Nucleolar localization signal (NoLS),Hypoxia, |
出版年 : | 2022 |
學位: | 碩士 |
摘要: | 肺結核(tuberculosis, TB)是由結核分枝桿菌(Mycobacterium tuberculosis, Mtb)感染所引起的疾病,在先前的研究中已知,受干擾素(interferons, INFs)調控表現的核蛋白SP110,可藉由調控先天免疫反應來對抗病原體的感染。此外,SP110的基因多態性(genetic polymorphisms)與宿主對肺結核的易感性相關。SP110蛋白是SP100/SP140蛋白家族中的一員,這類蛋白會在細胞核中與核小體(nuclear bodies, NBs)共定位(co-localization)形成聚集體。許多核小體蛋白在轉錄、細胞分化、細胞凋亡、老化和對DNA受損及感染的反應中扮演很重要的作用。在實驗室先前的研究中,確認在SP110蛋白的中間區域具有兩段核定位訊號(nuclear localization signal, NLS),且透過綠色螢光蛋白標定SP110275-314(NLS1)的蛋白定位表現,發現NLS1可能是一個潛藏的核仁定位訊號(nucleolar localization signal, NoLS)。然而雖然有核仁定位訊號的存在,表現在H1299肺癌細胞中的SP110b蛋白也只有很低的比例(小於5%)會使SP110b蛋白定位在核仁。實驗室近期的研究中發現,在微氧環境下SP110b蛋白會大幅轉移到核仁。且發現核仁定位訊號突變的SP110b Mut1+3突變蛋白,即使在常氧環境下也會有很高的比例定位在核仁。為了探索 NoLS 在 SP110 蛋白中的作用,本研究產生一系列突變 NoLS 的 SP110b 突變蛋白,並建立持續表現SP110b突變蛋白的H1299單一細胞株。接著分析NoLS的突變對SP110b在細胞中定位的影響,並進一步了解在面對微氧壓力時,細胞定位對H1299肺癌細胞生長的影響,以探討NoLS對SP110b功能的作用。 Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb) infection, and host genetics contribute to the infection outcomes. In our previous study, we reported that SP110 gene, which encodes an interferons (IFNs)-induced nuclear protein, is a genetic determinant that confers host innate immunity to Mtb infection. The SP110 protein is a member of SP100/SP140 protein family, and these proteins form aggregations that co-localize with nuclear bodies (NBs) in the nucleus. Many NB proteins have been revealed to play a key role in the regulation of transcription, cell division, apoptosis, senescence, and response to DNA damage or pathogen infection. Protein structural analyses showed that the SP110 contains several functional domains and may function as a transcriptional co-activator or co-repressor. Recently, two potential nuclear localization signals (NLSs) in the middle region of the SP110 protein have been identified. Besides, one of the NLSs, NLS1, has been demonstrated to direct the SP110 protein into the nucleolus, indicating that the NLS1 is a cryptic nucleolar localization signal (NoLS). Although there is an NoLS in SP110 protein, the protein localized to the nucleolus at a low frequency in H1299 lung cancer cell. However, our recent studies showed that the percentage of nucleolar-localized SP110 protein would significantly increase under hypoxia. To explore the role of the NoLS in the SP110 protein, SP110b mutants with a series of mutated NoLS were generated in this study. Their cellular localization as well as their effect on cell death were examined to clarify the role of the NoLS in the SP110 function. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79230 |
DOI: | 10.6342/NTU202210039 |
全文授權: | 同意授權(全球公開) |
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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