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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄧述諄(Shu-Chun Tung) | |
| dc.contributor.author | Cheng-Yen Chang | en |
| dc.contributor.author | 張正妍 | zh_TW |
| dc.date.accessioned | 2021-06-08T01:27:23Z | - |
| dc.date.copyright | 2015-03-12 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-07-30 | |
| dc.identifier.citation | 1. Conrad, M.N., et al., RAP1 protein interacts with yeast telomeres in vivo: overproduction alters telomere structure and decreases chromosome stability. Cell, 1990. 63(4): p. 739-50.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18804 | - |
| dc.description.abstract | 端粒是維持染色體末端穩定性重要的構造，在酵母菌中，Rap1為一DNA結合蛋白，能和有結合關係的蛋白質一起作用去保護端粒，防止染色體末端和另外一條染色體末端因為修復的關係而不正常的連結在一起(NHEJ)，並且保護端粒不會受到由端粒酶調控所引起不正常端粒延長的現象。在本篇研究中我們發現，在端粒縮短及DNA雙股斷裂下能引起Rap1蛋白質中絲胺酸731位點上發生磷酸化的現象，並且這個磷酸化是透過Mec1/Tel1所控制。我們也發現Rap1的磷酸化能保護端粒，防止端粒和端粒之間以直接黏接的方式形成連結的結構。除此之外，我們發現Rap1的磷酸化能夠促使Rap1和Rif2蛋白質停留在端粒上。總結以上，我們的發現暗示Rap1蛋白質的磷酸化對於端粒的調控是重要的。 | zh_TW |
| dc.description.abstract | Telomere maintenance is required for chromosome stability. In yeast Saccharomyces cerevisiae, the telomeres double strands binder, Rap1, together with interaction proteins protects telomeres from end-to-end fusion and telomere-mediated elongation. In this study, we found that under telomere shortening and double strand breaks, Rap1 serine 731 could be phosphorylated by Mec1/Tel1 check point kinases. Importantly, Rap1 phosphorylation could protect telomeres from non-homologous end join (NHEJ). Moreover, the phosphorylation could enhance Rap1 and Rif2 proteins binding on telomeres. All together, these results suggest that phosphorylation on Rap1 serves as an essential post-translational modification mark on telomere regulation. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T01:27:23Z (GMT). No. of bitstreams: 1 ntu-103-R01445112-1.pdf: 1440023 bytes, checksum: eb207cc8add7870df8c8d9c844884c45 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員會審定書………………………………………………………….……..……#
誌謝……………………………………………………………………...……………... 1 中文摘要……………………………………………………………………...………... 3 ABSTRACT…………………………………………………………………..………...4 CONTENTS…………………………………………………………………..………...5 Chapter 1 Introduction………………………………………………………………8 Chapter 2 Materials and Methods……………………………………………..…..13 2.1 Yeast Strains…………………………………………………………………...13 2.2 Southern Blot Analysis and Telomere Length Measurement………………….13 2.3 Antibodies and Western Blot Analysis………………………………………...13 2.4 Immunoprecipitation-Kinase Analysis………………………………………...14 2.5 Chromatin Immunoprecipitation……………………………………….…...…15 2.6 Amplification of the Telomere-Telomere Fusions by PCR……………………15 2.7 Telomeric Silencing Assays…………………………………………………....16 Chapter 3 Results………………………………………………………………...….17 3.1 Rap1pS731 Affects Telomere Length Maintenance…………………………...17 3.2 Phosphorylation of Rap1-S731 Depends on Mec1 and Tel1……………...…...18 3.3 Telomere Length Regulation of Phosphorylation of Rap1-S731 under MMS Treatment and yku80Δ cells…………………………………………………….19 3.4 Phosphorylation of Rap1-S731 Has Higher Rap1 and Rif2 Binding on Telomeres……………………………………………………………………….20 3.5 Phosphorylation of Rap1-S731 Prevents Telomere-telomere Fusions……...…21 3.6 Phosphorylation of Rap1-S731 Is Not Involved in Telomere Position Effect…22 3.7 Rap1 Phosphorylation Affects Both Rif1 and Rif2 Dependent Telomere Length Regulation……………………………………………………………………..23 Chapter 4 Discussion……………………………………………………………......25 4.1 The Phosphorylation of Rap1-S731 Is Dependent on Mec1 and Tell…………25 4.2 Phosphorylation of Rap1-S731 Exhibits Higher Rap1 Binding on Telomeres..27 4.3 Phosphorylation of Rap1-S731 Prevents Telomere-telomere Fusions…...……27 4.4 Phosphorylation of Rap1-S731 Shows No Direct Effect on TPE………..…....28 4.5 Rap1 Phosphorylation Affects Both Rif1 and Rif2 Dependent Telomere Length Regulation………………………………………………………………………29 4.6 Model for Phosphorylation of Rap1-S731 on Telomere Regulation…………..30 Chapter 5 References…………………………………………………………….....31 Chapter 6 Tables and Figures ….……………………………………………….…36 Table 1 Yeast Strains Used in This Study………………………………………….36 Table 2 Kinase Prediction by Protein Sequence…………………………………...39 Figure 1 Telomere Analysis of Rap1Phosphorylation Mutants...………………….40 Figure 2 Phosphorylation of Rap1-S731 Is Dependent on Mec1 and Tel1………..41 Figure 3 Telomere Length Regulation of Phosphorylation of Rap1-S731 under MMS Treatment and yku80Δ Cells………………………………………………......43 Figure 4 Phosphorylation of Rap1-S731 Presents Higher Rap1 and Rif2 Protein Bindings on Telomeres………………………………………………………………45 Figure 5 Phosphorylation at Rap1-S731 Could Suppress Telomere-telomere Fusions……………………………………………………………………………….47 Figure 6 Phosphorylation on Rap1-S731 Is Not related to Silencing Effect on Telomeres…………………………………………………………………………….48 Figure 7 Rap1 Phosphorylation Affects Both Rif1 and Rif2 Dependent Telomere Length Regulation…………………………………………………………………...50 Figure 8 Schematic Illustration of the Function of Rap1-S731 Phosphorylation on Telomeres……………………………………………………………...…………..…52 Figure S1 LC-Mass/Mass Analysis of Phosphorylation on Rap1…………………53 | |
| dc.language.iso | en | |
| dc.title | 探討Rap1蛋白質磷酸化在端粒的功能 | zh_TW |
| dc.title | Investigating the Functions of Phosphorylated Rap1 on Telomere Regulation | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林敬哲(Jing-Jer Lin),李財坤(Tsai-Kun Li) | |
| dc.subject.keyword | Rap1,磷酸化,NHEJ,端粒調控,Rif2,Mec1,Tel1, | zh_TW |
| dc.subject.keyword | Rap1,Phosphorylation,NHEJ,Telomere regulation,Rif2,Mec1,Tel1, | en |
| dc.relation.page | 54 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2014-07-30 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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