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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18265完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 董桂書(Kuei-Shu Tung) | |
| dc.contributor.author | Chia-Jou Lin | en |
| dc.contributor.author | 林佳柔 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:57:11Z | - |
| dc.date.copyright | 2020-09-17 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-08-19 | |
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Positioning of the Mitotic Spindle by a Cortical-Microtubule Capture Mechanism. Science, 287(5461), 2260. Li, X. C., Barringer, B. C., and Barbash, D. A. (2009). The pachytene checkpoint and its relationship to evolutionary patterns of polyploidization and hybrid sterility. 102(1), 24-30. Liu, Y.-C. (2016). The role of a spindle position checkpoint protein, Tem1, in yeast sporulation. Institute of Molecular and Cellular Biology. Taipei, Taiwan, National Taiwan University. 1-81. Maekawa, H., Priest, C., Lechner, J., Pereira, G., and Schiebel, E. (2007). The yeast centrosome translates the positional information of the anaphase spindle into a cell cycle signal. The Journal of cell biology, 179(3), 423-436. Maier, P., Rathfelder, N., Maeder, C. I., Colombelli, J., Stelzer, E. H. K., and Knop, M. (2008). The SpoMBe pathway drives membrane bending necessary for cytokinesis and spore formation in yeast meiosis. The EMBO journal, 27(18), 2363-2374. Markus, S. M., Kalutkiewicz, K. 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Ras recruits mitotic exit regulator Lte1 to the bud cortex in budding yeast. The Journal of cell biology, 161(5), 889-897. Zhou, B.-B. S., and Elledge, S. J. (2000). The DNA damage response: putting checkpoints in perspective. 408(6811), 433-439. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18265 | - |
| dc.description.abstract | 當環境缺乏養分時，會使二倍體出芽酵母菌細胞進行減數分裂並產生孢子。對於出芽酵母菌來說這是一個營養缺乏的逆境反應，因此會誘導出一些逆境蛋白，包括Hsp26蛋白。我們實驗室先前已經發現一個與Hsp26相關的紡錘絲檢控點，其會監控第二次減數分裂過程中的紡錘絲是否適合形成孢子。之前研究也發現Tem1參與在這個檢控點中。我們想了解Tem1是如何參與在此Hsp26相關紡錘絲檢控點。因為Tem1是GTP水解酶會受GAP複合物加速GTP進行水解，我們使用兩種方法將Tem1維持在GTP狀態並探討其效應。第一，是將GAP相關基因BFA1剔除以阻止GTP水解。第二，建構TEM1-Q79L突變株，組成持續與GTP結合的Tem1。我們發現若維持Tem1-GTP狀態，會使檢控點失去功能。透過pCLB2-TEM1突變細胞關閉減數分裂中Tem1表現也會使此一檢控點失效，據此我們提出兩種可能的模型來解釋此現象，一、Tem1-GDP可以主動阻擋的方式干擾孢子形成；二、Tem1必須存在才能活化Hsp26紡錘絲檢控點，接著此檢控點會反過來調控Tem1在孢子形成中的功能。 | zh_TW |
| dc.description.abstract | In the budding yeast Saccharomyces cerevisiae, absence of nitrogen and presence of nonfermentable carbon source lead diploid cells to enter meiosis and spore formation, which called sporulation. It is a starvation-stress response to the yeast. During sporulation, several stress proteins are induced, including Hsp26. Our laboratory has discovered that there is a Hsp26-dependent spindle checkpoint to monitor spindle assembly at meiosis II and regulates spore formation. Tem1, a component of the mitotic spindle checkpoint, is also involved in this checkpoint. We would like to know how Tem1 is involved in this Hsp26-dependent spindle checkpoint. Because Tem1 is a GTPase, which switches between GTP and GDP status, we adopted two approaches to keep Tem1 in the GTP-bound form. One was to delete the GAP-related gene, BFA1, and to block GTP hydrolysis. The other one was to construct TEM1-Q79L mutant that keeps Tem1 in the GTP-bound form. We found that Tem1-GTP may inactivates this checkpoint. pCLB2-TEM1 mutants, which shut down Tem1 during meiosis, also inactivate this checkpoint. We propose two models to explain these results. First, Tem1-GDP may actively block spore formation. Alternatively, Tem1 is required to activate the Hsp26-dependent spindle checkpoint that in turn regulates Tem1 function in spore formation. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:57:11Z (GMT). No. of bitstreams: 1 U0001-1308202010232400.pdf: 2228595 bytes, checksum: 10ab807f1f9e6eefd4d4151b2a6b4dbc (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 誌謝------------------------------------------------------------------------------------------------i 中文摘要-----------------------------------------------------------------------------------------ii ABSTRACT----------------------------------------------------------------------------------------iii TABLE OF CONTENTS--------------------------------------------------------------------------iv LIST OF TABLES----------------------------------------------------------------------------------viii LIST OF FIGURES--------------------------------------------------------------------------------x CHAPTER 1. INTRODUCTION--------------------------------------------------------------1 I. Sporulation---------------------------------------------------------------------------------1 A. Meiosis----------------------------------------------------------------------------------1 B. Spore formation-----------------------------------------------------------------------3 1. Formation of the meiotic II outer plaque------------------------------------3 2. Formation of the prospore membrane---------------------------------------4 (1) Septins---------------------------------------------------------------------------4 (2) Leading edge complex--------------------------------------------------------5 3. Prospore membrane closure----------------------------------------------------5 4. Spore wall assembly---------------------------------------------------------------6 II. Checkpoints--------------------------------------------------------------------------------6 A. Checkpoints in mitosis---------------------------------------------------------------7 1. DNA replication checkpoint------------------------------------------------------7 2. DNA damage checkpoint ---------------------------------------------------------8 3. Cell morphogenesis checkpoint-------------------------------------------------8 4. Spindle assembly checkpoint----------------------------------------------------8 5. Spindle position checkpoint------------------------------------------------------9 (1) Mitotic exit network-----------------------------------------------------------10 B. Checkpoints in meiosis---------------------------------------------------------------11 1. Pachytene checkpoint-------------------------------------------------------------11 2. Spindle assembly checkpoint----------------------------------------------------12 III. The meiotic Hsp26-dependent spindle checkpoint------------------------------13 A. The function of Hsp26 in sporulation --------------------------------------------13 1. Yeast Hsp26 protein----------------------------------------------------------------13 2. The research in Hsp26-dependent spindle checkpoint--------------------14 B. The small GTPase Tem1--------------------------------------------------------------15 1. Tem1 is a small G protein---------------------------------------------------------15 2. The universal switch mechanism-----------------------------------------------16 (1) Tem1 GTPase cycle-------------------------------------------------------------17 (2) Lte1 is long envisaged as a GEF for Tem1---------------------------------17 3. The localization of Tem1 and Bfa1/Bub2 in mitosis and meiosis--------18 VI. Specific Aims------------------------------------------------------------------------------19 CHAPTER 2. MATERIAL AND METHODS--------------------------------------------------20 I. Culture Conditions-------------------------------------------------------------------------20 II. DNA Preparation and Transformation------------------------------------------------21 III. Plasmids Constructions------------------------------------------------------------------22 IV. Yeast Strains Constructions-------------------------------------------------------------23 V. Benomyl Treatment------------------------------------------------------------------------26 VI. Spore Viability Analysis------------------------------------------------------------------27 CHAPTER 3. RESULTS--------------------------------------------------------------------------28 Analysis of sporulation in bfa1 mutants-------------------------------------------------28 Sporulation frequency in the bfa1 mutant cells was increased-----------------28 The bfa1 mutant was not affected with benomyl but fluctuate in sporulation --------------------------------------------------------------------------------------------------28 The sporulation defects caused by the kar9 mutation could be suppressed by the bfa1 mutation--------------------------------------------------------------------------30 Sporulation frequency in the bfa1hsp26 mutants----------------------------------32 GAP-related mutant strains enhance the sporulation frequency---------------33 Analysis of sporulation in TEM1-Q79L mutant-----------------------------------------34 Sporulation frequency in the TEM1-Q79L cells was increased------------------34 Benomyl does not affect sporulation in TEM1-Q79L cells------------------------35 Benomyl-treated TEM1-Q79L mutant cells displayed a decline in spore viability --------------------------------------------------------------------------------------------------36 TEM1-Q79L and pCLB2-TEM1 displayed similar phenotype in sporulation--37 Tem1-GTP mutants and pCLB2-TEM1 displayed similar phenotype in sporulation --------------------------------------------------------------------------------------------------38 CHAPTER 4. DISCUSSIONS--------------------------------------------------------------------40 The bfa1 mutant in sporulation------------------------------------------------------------40 The bfa1 mutation rescues the sporulation defect caused by kar9 mutation---41 The bfa1hsp26 mutant in sporulation-----------------------------------------------------42 The TEM1-Q79L mutant in sporulation---------------------------------------------------43 The contradiction between pCLB2-TEM1 and TEM1-Q79L mutants---------------44 (1) Tem1-GDP may actively block spore formation---------------------------------45 (2) Tem1 is required to activate the Hsp26-dependent spindle checkpoint that in turn regulates Tem1 function in spore formation-----------------------------------------46 REFERENCES----------------------------------------------------------------------------------------47 APPENDIX-------------------------------------------------------------------------------------------78 | |
| dc.language.iso | en | |
| dc.title | 過度活化的Tem1 GTPase對酵母菌孢子生成的影響 | zh_TW |
| dc.title | Studies of the Effect of Hyperactivated Tem1 GTPase on Yeast Sporulation | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡皇龍(Huang-Lung Tsai),王雅筠(Ya-Yun Wang) | |
| dc.subject.keyword | Hsp26,Tem1,Bfa1,GTP酶,GTP,紡錘絲檢控點,孢子形成過程, | zh_TW |
| dc.subject.keyword | Hsp26,Tem1,Bfa1,GTPase,GTP,spindle checkpoint,sporulation, | en |
| dc.relation.page | 78 | |
| dc.identifier.doi | 10.6342/NTU202003206 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2020-08-19 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子與細胞生物學研究所 | |
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