請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79230
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 顏伯勳 | zh_TW |
dc.contributor.advisor | Bo-Shiun Yan | en |
dc.contributor.author | 顏廷涓 | zh_TW |
dc.contributor.author | Ting-Chuan Yen | en |
dc.date.accessioned | 2022-11-17T17:01:12Z | - |
dc.date.available | 2023-11-09 | - |
dc.date.copyright | 2022-11-14 | - |
dc.date.issued | 2022 | - |
dc.date.submitted | 2022-11-10 | - |
dc.identifier.citation | 1. World Health Organization. Global tuberculosis report 2022. [https://www.who.int/news-room/fact-sheets/detail/tuberculosis] 2022.
2. Centers for Disease Control and Prevention Tuberculosis (TB). 2022; Available from: https://www.cdc.gov/tb/default.htm. 3. F.O. Vannberg, Chapman, S.J., and Hill, A.V., Human genetic susceptibility to intracellular pathogens. Immunol Rev, 2011. 240(1): p. 105-16, doi: 10.1111/j.1600-065X.2010.00996.x. 4. J.L. Casanova and Abel, L., Genetic dissection of immunity to mycobacteria: the human model. Annu Rev Immunol, 2002. 20: p. 581-620, doi: 10.1146/annurev.immunol.20.081501.125851. 5. A.V. Hill, The genomics and genetics of human infectious disease susceptibility. Annu Rev Genomics Hum Genet, 2001. 2: p. 373-400, doi: 10.1146/annurev.genom.2.1.373. 6. C.J. Lynch, Pierce-Chase, C.H., and Dubos, R., A GENETIC STUDY OF SUSCEPTIBILITY TO EXPERIMENTAL TUBERCULOSIS IN MICE INFECTED WITH MAMMALIAN TUBERCLE BACILLI. J Exp Med, 1965. 121(6): p. 1051-70, doi: 10.1084/jem.121.6.1051. 7. I. Kramnik, Genetic dissection of host resistance to Mycobacterium tuberculosis: the sst1 locus and the Ipr1 gene. Curr Top Microbiol Immunol, 2008. 321: p. 123-48, doi: 10.1007/978-3-540-75203-5_6. 8. H. Pan, et al., Ipr1 gene mediates innate immunity to tuberculosis. Nature, 2005. 434(7034): p. 767-72, doi: 10.1038/nature03419. 9. R. Bernardi and Pandolfi, P.P., Structure, dynamics and functions of promyelocytic leukaemia nuclear bodies. Nat Rev Mol Cell Biol, 2007. 8(12): p. 1006-16, doi: 10.1038/nrm2277. 10. V. Lallemand-Breitenbach and de Thé, H., PML nuclear bodies. Cold Spring Harb Perspect Biol, 2010. 2(5): p. a000661, doi: 10.1101/cshperspect.a000661. 11. D.B. Bloch, et al., Sp110 localizes to the PML-Sp100 nuclear body and may function as a nuclear hormone receptor transcriptional coactivator. Mol Cell Biol, 2000. 20(16): p. 6138-46, doi: 10.1128/mcb.20.16.6138-6146.2000. 12. J. Pitkänen and Peterson, P., Autoimmune regulator: from loss of function to autoimmunity. Genes Immun, 2003. 4(1): p. 12-21, doi: 10.1038/sj.gene.6363929. 13. S. Kadereit, et al., Molecular cloning of two new interferon-induced, highly related nuclear phosphoproteins. J Biol Chem, 1993. 268(32): p. 24432-41, doi: 14. G.J. Fox, et al., Polymorphisms of SP110 are associated with both pulmonary and extra-pulmonary tuberculosis among the Vietnamese. PLoS One, 2014. 9(7): p. e99496, doi: 10.1371/journal.pone.0099496. 15. J.S. Leu, et al., SP110b Controls Host Immunity and Susceptibility to Tuberculosis. Am J Respir Crit Care Med, 2017. 195(3): p. 369-382, doi: 10.1164/rccm.201601-0103OC. 16. J.S. Szeszko, et al., Resequencing and association analysis of the SP110 gene in adult pulmonary tuberculosis. Hum Genet, 2007. 121(2): p. 155-60, doi: 10.1007/s00439-006-0293-z. 17. K. Tosh, et al., Variants in the SP110 gene are associated with genetic susceptibility to tuberculosis in West Africa. Proc Natl Acad Sci U S A, 2006. 103(27): p. 10364-10368, doi: 10.1073/pnas.0603340103. 18. J.-S. Leu, et al., Functional domains of SP110 that modulate its transcriptional regulatory function and cellular translocation. Journal of Biomedical Science, 2018. 25(1): p. 34, doi: 10.1186/s12929-018-0434-4. 19. J. Lu, et al., Types of nuclear localization signals and mechanisms of protein import into the nucleus. Cell Communication and Signaling, 2021. 19(1): p. 60, doi: 10.1186/s12964-021-00741-y. 20. L. Stenström, et al., Mapping the nucleolar proteome reveals a spatiotemporal organization related to intrinsic protein disorder. Mol Syst Biol, 2020. 16(8): p. e9469, doi: 10.15252/msb.20209469. 21. T. Miyake and McDermott, J.C., Nucleolar localization of c-Jun. Febs j, 2022. 289(3): p. 748-765, doi: 10.1111/febs.16187. 22. D. Hernandez-Verdun, et al., The nucleolus: structure/function relationship in RNA metabolism. Wiley Interdiscip Rev RNA, 2010. 1(3): p. 415-31, doi: 10.1002/wrna.39. 23. A. Di Bacco, et al., The SUMO-specific protease SENP5 is required for cell division. Mol Cell Biol, 2006. 26(12): p. 4489-98, doi: 10.1128/mcb.02301-05. 24. A.K. Leung, et al., Quantitative kinetic analysis of nucleolar breakdown and reassembly during mitosis in live human cells. J Cell Biol, 2004. 166(6): p. 787-800, doi: 10.1083/jcb.200405013. 25. I. Grummt, The nucleolus—guardian of cellular homeostasis and genome integrity. Chromosoma, 2013. 122(6): p. 487-97, doi: 10.1007/s00412-013-0430-0. 26. L. Hua, et al., Nucleolus and Nucleolar Stress: From Cell Fate Decision to Disease Development. Cells, 2022. 11(19), doi: 10.3390/cells11193017. 27. K. Peltonen, et al., Small molecule BMH-compounds that inhibit RNA polymerase I and cause nucleolar stress. Mol Cancer Ther, 2014. 13(11): p. 2537-46, doi: 10.1158/1535-7163.Mct-14-0256. 28. A. Michienzi, et al., A nucleolar localizing Rev binding element inhibits HIV replication. AIDS Res Ther, 2006. 3: p. 13, doi: 10.1186/1742-6405-3-13. 29. J. Lee, et al., Nucleolar dysfunction in Huntington's disease. Biochim Biophys Acta, 2014. 1842(6): p. 785-90, doi: 10.1016/j.bbadis.2013.09.017. 30. R. Parlato and Kreiner, G., Nucleolar activity in neurodegenerative diseases: a missing piece of the puzzle? Journal of Molecular Medicine, 2013. 91(5): p. 541-547, doi: 10.1007/s00109-012-0981-1. 31. C.-P. Yang, et al., Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58). Journal of Biomedical Science, 2015. 22(1): p. 33, doi: 10.1186/s12929-015-0136-0. 32. A. Emami Nejad, et al., The role of hypoxia in the tumor microenvironment and development of cancer stem cell: a novel approach to developing treatment. Cancer Cell International, 2021. 21(1): p. 62, doi: 10.1186/s12935-020-01719-5. 33. M.A.E. Lohrum, et al., Identification of a cryptic nucleolar-localization signal in MDM2. Nature Cell Biology, 2000. 2(3): p. 179-181, doi: 10.1038/35004057. | - |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79230 | - |
dc.description.abstract | 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。 | zh_TW |
dc.description.abstract | 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. | en |
dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2022-11-17T17:01:12Z
No. of bitstreams: 0 | en |
dc.description.provenance | Made available in DSpace on 2022-11-17T17:01:12Z (GMT). No. of bitstreams: 0 | en |
dc.description.tableofcontents | 誌謝 i
摘要 ii Abstract iii 圖目錄 vii 附錄 viii Chapter 1 緒論 1 1.1 肺結核(Tuberculosis, TB) 1 1.2 SP110 2 1.3 核定位訊號(nuclear localization signal, NLS 3 1.4 核仁(nucleolus) 3 1.5 核仁定位訊號(nucleolar localization signal, NoLS)4 1.6 微氧環境(hypoxia)5 1.7 研究目標 5 Chapter 2 實驗材料與方法 6 2.1 細胞株和細胞培養 6 2.2 質體 (Plasmids) 6 2.3 細胞轉染(Transfection)7 2.4 慢病毒生產(Lentivirus production)7 2.5 慢病毒轉導(Lentivirus transduction) 7 2.6 單細胞克隆(Single clones selection)8 2.7 西方墨點法(Western blot) 8 2.8 免疫螢光染色(Immunofluorescence staining) 8 2.9 細胞群落形成分析(Colony formation assay, CFA) 9 2.10 細胞存活率分析(MTT assay) 9 2.11 統計分析 10 Chapter 3 結果 11 3.1 建構SP110b突變株質體 11 3.2 確認細胞中SP110b蛋白表現 12 3.3 建立SP110b突變體單一細胞株 12 3.4 NoLS突變影響SP110b定位在核仁的比例 13 3.5 SP110b定位影響H1299細胞群落生長 14 3.6 SP110b定位影響H1299細胞生長趨勢 15 Chapter 4 結論與討論 16 Chapter 5 參考資料 19 Chapter 6 圖附錄 23 Chapter 7 附錄 45 | - |
dc.language.iso | zh_TW | - |
dc.title | 一段潛藏的核仁定位訊號在SP110蛋白功能的作用 | zh_TW |
dc.title | The role of a cryptic nucleolar-localization signal in SP110 protein function | en |
dc.title.alternative | The role of a cryptic nucleolar-localization signal in SP110 protein function | - |
dc.type | Thesis | - |
dc.date.schoolyear | 111-1 | - |
dc.description.degree | 碩士 | - |
dc.contributor.oralexamcommittee | 詹世鵬;蘇剛毅 | zh_TW |
dc.contributor.oralexamcommittee | Shih-Peng Chan;Kang-Yi Su | en |
dc.subject.keyword | SP110,細胞定位,核定位訊號,核仁定位訊號,微氧壓力, | zh_TW |
dc.subject.keyword | SP110,Cellular localization,Nuclear localization signal (NLS),Nucleolar localization signal (NoLS),Hypoxia, | en |
dc.relation.page | 50 | - |
dc.identifier.doi | 10.6342/NTU202210039 | - |
dc.rights.note | 同意授權(全球公開) | - |
dc.date.accepted | 2022-11-10 | - |
dc.contributor.author-college | 醫學院 | - |
dc.contributor.author-dept | 生物化學暨分子生物學研究所 | - |
顯示於系所單位: | 生物化學暨分子生物學科研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
U0001-0619221109502016.pdf | 3.76 MB | Adobe PDF | 檢視/開啟 |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。