跳到主要內容

臺灣博碩士論文加值系統

(3.231.230.177) 您好!臺灣時間:2021/07/27 10:38
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:蔡岑儀
研究生(外文):Tsen-Yi Tsai
論文名稱:探討酵母菌中HTL1在RSC複合物中的特徵
論文名稱(外文):Characterization of yeast HTL1 containing RSC complex
指導教授:鄭明媛
指導教授(外文):Ming-Yuan Cheng
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:酵母菌
外文關鍵詞:yeast
相關次數:
  • 被引用被引用:0
  • 點閱點閱:94
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
目前已知RSC複合物對於基因和細胞週期調控上扮演著舉足輕重的角色。而越來越多的研究資料顯示,HTL1基因和RSC複合物的構型、種類和功能有密切的關係。在本論文中利用了離子交換層析法、蔗糖梯度分離、高拷貝抑制基因策略篩選和蛋白剪切酵素的處理,希望能更深入地了解HTL1的功能以及它和RSC複合物之間的交互作用。
在離子交換層析法和蔗糖梯度分離實驗中得知,在野生型菌株和htl1刪除株中,RSC複合物的成員是否有被DNase處理過所純化出來以及在蔗糖梯度溶液中的分佈不同。除此之外,也利用了蛋白剪切酵素的處理觀察到缺乏Htl1p時,Rsc8p對蛋白剪切酵素endopeptidase Glu-C較敏感,這可能就是缺乏Htl1p會影響到RSC複合物穩定性的原因。為了更進一步探討HTL1基因的特性,利用高拷貝抑制基因策略篩選抑制htl1刪除株對benomyl過度敏感的基因,結果沒有挑到抑制者。
目前大家認為htl1刪除是直接造成酵母菌高溫致死的原因。根據我們實驗室先前利用蔗糖濃度梯度的方式來分析比較在37℃下生長的野生型菌株 (SEY6211) 和htl1刪除 (CKLa2),發現缺乏Htl1p會影響RSC複合物的穩定性,使Rsc8p蛋白質和Sth1p蛋白質間的交互作用變的不穩定,然而,可能RSC複合物中的組成蛋白質間的交互作用不穩定才是造成酵母菌高溫致死的直接因素。為了更進一步觀察是否Rsc8p蛋白質和Sth1p蛋白質間的交互作用的穩定度是造成高溫致死現象的重要關鍵,因此設計了一個融合蛋白的實驗。我們將所建構出的STH1-LINKER-RSC8 (SLR) DNA序列轉殖入htl1刪除株中。目前觀察到有孢子可以生長在30℃和 37℃ SD-U培養基上。
RSC complex, an essential chromatin-remodeling complex in Saccharomyces cerevisiae, performs central roles in transcriptional regulation and cell cycle progression. RSC complex has been found to interact with Htl1p. To study the relationship between Htl1p and RSC complex, a series of analysis, including ion exchange column chromatography, sucrose gradient assay, multiple copy suppressor and partial protease, were performed in this thesis. By ion exchange column chromatography and sucrose gradient assay, the eluted protein and the distribution of subunits of RSC complex with or without DNase treatment were different between htl1 mutant and wild type cells. According to the analysis of sucrose gradient distribution in wild type and htl1 deletion strains at 37℃, we found Htl1p may play a role in maintaining stability of RSC complex, especially in the interaction between Rsc8p and Sth1p. Therefore, the instability of RSC complexes might be the main reason that causes temperature sensitivity. To further characterize if the interaction of Sth1p and Rsc8p is crucial for the temperature sensitive phenotype. A fusion protein experiment was designed. We constructed STH1-LINKER-RSC8 (SLR) fusion gene and inserted it into the strain without HTL1. So far that Ura+ spore can grow at 30℃ and 37℃. Partial protease digestion demonstrated that Rsc8p exhibits endopeptidase Glu-C sensitivity in htl1 deletion mutant. In order to further characterize HTL1 gene, we screened suppressors of Benomyl sensitivity of htl1 deletion strain by multicopy suppressor scheme. However, no suppressor of htl1 deletion strain was obtained.
參考文獻
1. Martin J. Romeo, Melinda L. Angus-Hill, Andrew K. Sobering, Yoshiaki Kamada, Bradley R. Cairns, and David E. Levin. (2002). HTL1 encodes a novel factor that interacts with the RSC chromatin-remodeling complex in Saccharomyces cerevisiae. Mol Cell Biol. 22(23):8165–8174.
2. Becker, P. B., and Horz, W. (2002). ATP-dependent chromatin remodeling. Annu. Rev. biochem. 70:247-273.
3. Boyer, L. A., Langer, M. R., Crowley, S. T., Denu, J. M., and Peterson, C. L. (2002). Essential role for the SANT domain in the functioning of multiple chromatin remodeling enzymes. Mol. Cell. 10:935-947.
4. Owen, D.G., Ornaghi, P., Yang, J.C., Lowe, N.,Evans, P. R., Ballario, P., Neuhaus, D., Filetic, P., and Traver, A. A. (2002). The structure basis for the recognition of acetylated and histone H4 by the bromodomain of histone acetyltransferase gcn5p. EMBO J. 19:6141-6149.
5. Lemmon, M. A., Ferguson, K. M., and Schlessinger, J. (1996). PH domain: diverse sequences with a common fold recruit signaling molecules to the cell surface. Cell 85:621-624.
6. Narlikar, G. J., Fan, H., and Kindston, R. E. (2002). Cooperation between complexes that regulate chromatin structure and transcription. Cell 108:475-487.
7. Langst, G., and Becker, P. B. (2001). Nucleosome mobilization and positioning by ISWI-containing chromatin-remodeling factor. J. Cell Sci. 114:2561-2568.
8. Afalf, J. D., Narilkar, G. J., and Kingston, R. E. (2000). Function differences between the human ATP-dependent nucleosome remodeling protein BRGI and SNF2H. J. Biol. Chem. 276:34270-34278.
9. Kingston, R. E., and Narlikar, G. J. (1999). ATP-dependent remodeling and acetylation as regulators of chromatin fluidity. Gene Dev. 13:2339-2352.
10. Cairns BR, Erdjument-Bromage H, Tempst P, Winston F, Kornberg RD. (1998). Two actin-related proteins are shared functional components of the components of the chromatin-remodeling complex RSC and SWI/SNF. Mol. Cell. 2:639-659.
11. Cairns BR, Schlichter A, Erdjument-Bromage H, Tempst P, Kornberg RD, Winston F. (1999). Two functionally distinct forms of the RSC nucleosome remodeling complex, containing essential AT hook, BAH, and bromodomains. Mol. Cell. 4(5):715-23.
12. Lorch Y, Zhang M, and Kornberg RD. (1999). Histone octamer transfer by a chromatin-remodeling complex. Cell 96(3):389-92.
13. Cairns BR, Lorch Y, Li Y, Zhang M, Lacomis L, Erdjument-Bromage H, Tempst P, Du J, Laurent B, Kornberg R. (1996). RSC, an essential, abundant chrimatin-remodeling complex. Cell 87(7):1249-60.
14. Lorch Y, Cairns BR, Zhang M, and Kornberg RD. (1998). Activated RSC-nucleosome complex and persistently altered form of the nucleosome. Cell 94(1):29-34.
15. Hirofumi Koyama, Taka-aki Nagao, Tomomi Inai, Kohji Miyahara, Yasufumi Hayasida, Katsuhiko Shirahige, and Eiko Tsuchiya. (2004). RSC Nucleosome-remodeling Complex Plays Prominent Roles in Transcriptional Regulation throughout Budding Yeast Gametogenesis. Boisci. Biotechnol. Biochem. 68(4):909- 919.
16. Melinda L. A., Alisha S., Douglas R., Hediye E. B., Paul T. and Bradley R. C. (2001). A RSC3/RSC30 Zinc cluster dimmer reveals novel roles for the chromatin remodeler RSC in gene expression and cell cycle control. Mol. Cell. 7:741-751.
17. Margaret Kasten1, Heather Szerlong1, Hediye Erdjument-Bromage2, Paul Tempst2, Michel Werner3 and Bradley R Cairns1. (2004.) Tandem bromodomains in the chromatin remodeler RSC recognize acetylated histone H3 Lys14. The EMBO Journal. 23:1348–1359.
18. 黃姿雯。(2000)。功能性分析酵母菌SHL1基因。陽明大學,遺傳學研究所碩士班論文。
19. 楊孟鈴。(2001)。功能性分析酵母菌SHL1基因。陽明大學,生命科學學士論文。
20. T. Taneda, A. Kikuchi. (2004). Genetic Analysis of RSC58, Which encodes a component of a yeast chromatin remodeling complex, and interacts with the transcription factor Swi6. Mol Genet Genomics. 271: 479-489.
21. 蔡艾莉。(2002)。探討酵母菌Htl1p與RSC複合物之關係。陽明大學,遺傳學研究所碩士班論文。
22. Bender A, Pringle JR. (1991). Use of a screen for synthetic lethal and multicopy suppressor mutants to identify two new genes involved in morphogenesis in Saccharomyces cerevisiae. Mol Cell Biol. 11(3):1295-305.
23. Hosotani T, Koyama H, Uchino M, Miyakawa T, Tsuchiya E. (2001). PKC1, a protein kinase C homologue of Saccharomyces cerevisiae, participates in microtubule function through the yeast EB1 homologue, BIM1. Genes Cells. 6:775-788.
24. Joseph A Martens and Fred Winston. (2003). Recent advances in understanding chromatin remodeling by Swi/Snf complexes. Current Opinion in Genetics & Development. 13:136–142.
25. Ng HH, Robert F, Young RA, Struhl K. (2002). Genome-wide location and regulated recruitment of the RSC nucleosome-remodeling complex. Genes Dev. 16:806-819.
26. Damelin, M., Simon, I., Moy, T. I., Wilson, S., Tempst, P., Roth, F. P., Young, R. A., Cairns, B. R., and Silver, P. A. (2002). The genome-wide localization of Rsc9, a component of the RSC chromatin-remodeling complex, changes in response to stress. Molecular Cell. 9:563-573.
27. Lanzuolo, C., S. Ederle, A., Pollice, F. Russo, A. Storlazzi, and J. F. Pulitzer. (2001). The HTL1 gene (YCR020W-b) of Saccharomyces cerevisiae is necessary for growth at 37 degrees C, and for the conservation of chromosome stability and fertility. Yeast 18:1317-30.
28. 張緯倫。(2001)。探討酵母菌Htl1p與RSC複合物之關係。陽明大學,生命科學系學士論文。
29. Carlson, M., and Botstein, D. (1982). Two differentially regulated mRNA with different 5’ ends encode secreted and intracellular forms of yeast invertase. Cell. 28:145-154.
30. Lu YM, Lin YR, Tsai A, Hsao YS, Li CC, Cheng MY. (2003). Dissecting the pet18 mutation in Saccharomyces cerevisiae: HTL1 encodes a 7-kDa polypeptide that interacts with components of the RSC complex. Mol Genet Genomics. 269(3):321-30.
31. Praross-Macdonald, Paulo S. R. Coelho, Terry Roemer, Seema Agarwal, Anuj Kumar, Ronald Jansen, Ken-Hoi Cheung, Amy Sheehan, Dawn Syoniatis. (1999). Large-scale analysis of the yeast genome by transposon tagging and gene disruption. Nature 402:413 – 418.
32. 王善立。(2003)。探討酵母菌Htl1p與RSC複合物之關係。陽明大學,遺傳學研究所碩士班論文。
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
1. 李銘義:「兩岸政治對話之美國因素」,《中國事務季刊》,第六期(2001年10月):頁92∼120。
2. 陳一新:「從布希外交團隊與領導風格看美國外交與兩岸政策」,《遠景季刊》,第二卷,第一期(2001年1月):頁1∼33。
3. 關中:「後冷戰時代美國外交政策的批判」,《遠景季刊》,第三卷,第一期(2002年1月):頁1∼26。
4. 虞義輝:「現階段中共對台統戰策略之回顧與評析」,《共黨問題研究》,第25卷,第5期,(民國88年5月),頁5∼18。
5. 于有慧:「中共外交政策走向與選擇」,《問題與研究》,第43卷,第1期(民國93年1、2月):頁105∼124。
6. 宋鎮照:「解析當前美『中』兩國亞太政策下的東南亞發展策略」,《展望與探索》,第1卷,第7期(民國92年7月):頁12∼27。
7. 蔡裕明:「後九一一時期中共外交政策的持續與遞嬗」,《共黨問題研究》,第28卷,第10期(民國91年10月):頁52~60。
8. 林正義:「布希政府台海兩岸政策初探」,《中國事務季刊》,第七期(2002年1月):頁67∼81。
9. 傅亞蘭:「九一一事件對美中台三邊關係的影響」,《共黨問題研究》,第28卷,第1期(民國91年1月):頁80∼83。
10. 莫大華&陳紫財:「美國介入未來台海危機影響因素分析之研究」,《共黨問題研究》,第二十八卷,第一期(民國91年1月):頁4∼16。
11. 秦邦九:「中共推動『多極化』世界格局與『夥伴外交』之研究」,《共黨問題研究》,第25卷,第9期(民國88年9月):頁40∼47。
12. 張錫模:「九一一事件後的中亞情勢」,《國際事務季刊》,第2卷(民國91年):頁82∼101。
13. 以理:「中共對美發展『戰區導彈防禦系統』(TMD)反應之研析」,《中共研究》,第34卷,第7期(2002年7月):頁70∼78。
14. 陳一新:「布希政府亞太戰略的構想與部署」,《遠景季刊》,第二卷,第四期(2001年10月):頁1∼27。
15. 王崑義:「美國的反恐佈主義與國際安全—兼論九一一事件以後台海兩岸的處境」,《遠景季刊》,第三卷,第二期,(2002年4月),頁137∼182。