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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 呂勝春 | |
dc.contributor.author | Yu-Sheng Lin | en |
dc.contributor.author | 林祐生 | zh_TW |
dc.date.accessioned | 2021-06-15T00:43:21Z | - |
dc.date.available | 2008-09-25 | |
dc.date.copyright | 2008-09-25 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-08-29 | |
dc.identifier.citation | 1. Cammas, F., et al., Mice lacking the transcriptional corepressor TIF1beta are defective in early postimplantation development. Development, 2000. 127(13): p. 2955-63.
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Ziv, Y., et al., Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway. Nat Cell Biol, 2006. 8(8): p. 870-6. 18. Eissenberg, J.C., Decisive factors: a transcription activator can overcome heterochromatin silencing. Bioessays, 2001. 23(9): p. 767-71. 19. Brasher, S.V., et al., The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. EMBO J, 2000. 19(7): p. 1587-97. 20. Wang, G., et al., Conservation of heterochromatin protein 1 function. Mol Cell Biol, 2000. 20(18): p. 6970-83. 21. Eissenberg, J.C. and S.C. Elgin, The HP1 protein family: getting a grip on chromatin. Curr Opin Genet Dev, 2000. 10(2): p. 204-10. 22. Nielsen, A.L., et al., Selective interaction between the chromatin-remodeling factor BRG1 and the heterochromatin-associated protein HP1alpha. EMBO J, 2002. 21(21): p. 5797-806. 23. 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Underhill, C., et al., A novel nuclear receptor corepressor complex, N-CoR, contains components of the mammalian SWI/SNF complex and the corepressor KAP-1. J Biol Chem, 2000. 275(51): p. 40463-70. 29. Ayyanathan, K., et al., Regulated recruitment of HP1 to a euchromatic gene induces mitotically heritable, epigenetic gene silencing: a mammalian cell culture model of gene variegation. Genes Dev, 2003. 17(15): p. 1855-69. 30. Peters, A.H., et al., Partitioning and plasticity of repressive histone methylation states in mammalian chromatin. Mol Cell, 2003. 12(6): p. 1577-89. 31. Stewart, M.D., J. Li, and J. Wong, Relationship between histone H3 lysine 9 methylation, transcription repression, and heterochromatin protein 1 recruitment. Mol Cell Biol, 2005. 25(7): p. 2525-38. 32. 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Cammas, F., et al., Association of the transcriptional corepressor TIF1beta with heterochromatin protein 1 (HP1): an essential role for progression through differentiation. Genes Dev, 2004. 18(17): p. 2147-60. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42039 | - |
dc.description.abstract | TIF1b 是一個上位遺傳調控因子,藉由與其它的染色質蛋白互相作用,能夠在基因體的特定區位上進行染色質的結構重整與轉錄調控。一般認為TIF1b 的抑制功能主要經由其與異染色質蛋白一號(Heterochromatin Protein 1, HP-1)的交互作用所達成。在TIF1b 上的HP-1 box 是主要負責這個交互作用的區段。然而,背後的調控機制仍未完全明瞭。在這篇論文中,我們建立了一個基因置換系統能夠將
內生性的蛋白質抑制,以外生性的突變蛋白質補回,藉以研究特定胺基酸位點對其功能之影響。此外,利用抗體的專一性,我們也發現TIF1b 能夠在HP-1 box 中的Ser473 上被磷酸化,而這樣的磷酸化是會隨著細胞週期而改變的。此外,利用染色質沉澱的技術,我們也觀察到這樣的磷酸化能夠確實影響到特定基因體區位上的HP-1 召集。最後,我們也觀察到了TIF1b 能夠與組蛋白乙烯基轉化脢PCAF形成錯合物,而此觀察也提供了TIF1b 做為活化因子的可能性。 | zh_TW |
dc.description.abstract | The transcriptional intermediary factor 1β (TIF1β)/KRIP-1/KAP-1/TRIM28 is an epigenetic regulator correlated to transcriptional regulation and chromatin remodeling at designated genomic loci through the interaction with other chromatinic proteins. The repressive capability of TIF1β is thought to be mediated in part by its interaction with HP1. The HP1-box, PXVXL, of TIF1β is reported to be responsible for this interaction.
However, the underlying regulation is still poorly understood. Here we demonstrate the construction of a gene replacement system which replaces the endogenous TIF1β with ectopically expressed ones carrying desired alterations. Also, using phospho-specific antibody, it is shown that TIF1β is phosphorylated at Ser473, which is located in the HP1-interacting domain, and the global level of this modification fluctuates with the progression of cell cycle. Furthermore, in our ChIP assay, it is shown that the overexpression of wild type and S473A but not the phospho-mimicry S473E mutant of TIF1β can preferentially enrich the recruitment of HP1β to endogenous promoter region of CDC2 and CDC25a. In conclusion, the phosphorylation of TIF1β Ser473 is demonstrated to be functionally correlated to the recruitment of HP1 protein to specific chromatinic loci. Finally, we’ve also provided the evidence that TIF1β can potentially complex with the histone acetyltransferases PCAF, and this interaction is possibly involved in the TIF1β-mediated transcriptional activation reported previously. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T00:43:21Z (GMT). No. of bitstreams: 1 ntu-97-R94448008-1.pdf: 612217 bytes, checksum: ea5622a55410dc5da43604152ce46161 (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 致謝 3
中文摘要 5 Abstract 6 Material and Methods 10 Cell Culture 10 Plasmid Construction 10 Transfection 11 Preparation of Whole Cell Extract 11 Cell Synchronization 11 Antibody 12 SDS-PAGE and Western Blot 12 Chromatin Immunoprecipitation 13 Immunostaining 14 Result 15 Construction of Vector-based miRNA against mouse TIF1β 16 TIF1β Replacement 16 The Dynamic Regulation of TIF1β Ser473 Phosphorylation during Cell Cycle Progression 16 Phospho-mimicry Mutation of TIF1β Ser473 Compromises the Recruitment of HP1β to E2F-responsive elements in vivo 17 Endogenous TIF1β can be Co-immunoprecipitated by Overexpressed PCAF 18 Discussion 20 Figure 23 Figure Legent 30 Reference 32 | |
dc.language.iso | en | |
dc.title | TIF1beta對細胞週期調控之功能分析 | zh_TW |
dc.title | The Functional Analysis of TIF1beta on the Regulation of Cell Cycle | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張?仁,譚賢明 | |
dc.subject.keyword | 磷酸化,轉錄調控, | zh_TW |
dc.subject.keyword | TIF1beta,HP-1,Epigenetics,Phosphorylation,Transcriptional Regulation, | en |
dc.relation.page | 35 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-08-29 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
顯示於系所單位: | 分子醫學研究所 |
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