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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄧述諄(Shu-Chun Teng) | |
| dc.contributor.author | Jia-Cin Jhang | en |
| dc.contributor.author | 張嘉琴 | zh_TW |
| dc.date.accessioned | 2021-06-15T05:08:56Z | - |
| dc.date.available | 2020-07-23 | |
| dc.date.copyright | 2010-09-09 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-07-26 | |
| dc.identifier.citation | [1] Bailly, E., S. Cabantous, et al. (2003). 'Differential cellular localization among mitotic cyclins from Saccharomyces cerevisiae: a new role for the axial budding protein Bud3 in targeting Clb2 to the mother-bud neck.' J Cell Sci 116(Pt 20): 4119-4130.
[2] Bloom, J. and F. R. Cross (2007). 'Multiple levels of cyclin specificity in cell-cycle control.' Nat Rev Mol Cell Biol 8(2): 149-160. [3] Carroll, C. W., R. Altman, et al. (1998). 'The septins are required for the mitosis-specific activation of the Gin4 kinase.' J Cell Biol 143(3): 709-717. [4] Chant, J. and I. Herskowitz (1991). 'Genetic control of bud site selection in yeast by a set of gene products that constitute a morphogenetic pathway.' Cell 65(7): 1203-1212. [5] Chant, J., M. Mischke, et al. (1995). 'Role of Bud3p in producing the axial budding pattern of yeast.' J Cell Biol 129(3): 767-778. [6] Drubin, D. G. (1991). 'Development of cell polarity in budding yeast.' Cell 65(7): 1093-1096. [7] Fujita, A., C. Oka, et al. (1994). 'A yeast gene necessary for bud-site selection encodes a protein similar to insulin-degrading enzymes.' Nature 372(6506): 567-570. [8] Gruhler, A., J. V. Olsen, et al. (2005). 'Quantitative phosphoproteomics applied to the yeast pheromone signaling pathway.' Mol Cell Proteomics 4(3): 310-327. [9] Hofken, T. and E. Schiebel (2002). 'A role for cell polarity proteins in mitotic exit.' EMBO J 21(18): 4851-4862. [10] Holt, L. J., B. B. Tuch, et al. (2009). 'Global analysis of Cdk1 substrate phosphorylation sites provides insights into evolution.' Science 325(5948): 1682-1686. [11] Justa-Schuch, D., Y. Heilig, et al. (2010). 'Septum formation is regulated by the RHO4-specific exchange factors BUD3 and RGF3 and by the landmark protein BUD4 in Neurospora crassa.' Mol Microbiol 76(1): 220-235. [12] Lew, D. J. and S. I. Reed (1993). 'Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins.' J Cell Biol 120(6): 1305-1320. [13] Lew, D. J. and S. I. Reed (1995). 'A cell cycle checkpoint monitors cell morphogenesis in budding yeast.' J Cell Biol 129(3): 739-749. [14] Lew, D. J. and S. I. Reed (1995). 'Cell cycle control of morphogenesis in budding yeast.' Curr Opin Genet Dev 5(1): 17-23. [15] Madden, K. and M. Snyder (1998). 'Cell polarity and morphogenesis in budding yeast.' Annu Rev Microbiol 52: 687-744. [16] McCusker, D., C. Denison, et al. (2007). 'Cdk1 coordinates cell-surface growth with the cell cycle.' Nat Cell Biol 9(5): 506-515. [17] Neuwald, A. F., J. S. Liu, et al. (1997). 'Extracting protein alignment models from the sequence database.' N ucleic Acids Res 25(9): 1665-1677. [18] Nigg, E. A. (2001). 'Mitotic kinases as regulators of cell division and its checkpoints.' Nat Rev Mol Cell Biol 2(1): 21-32. [19] Padmashree, C. G. and U. Surana (2001). 'Cdc28-Clb mitotic kinase negatively regulates bud site assembly in the budding yeast.' J Cell Sci 114(Pt 1): 207-218. [20] Park, H. O. and E. Bi (2007). 'Central roles of small GTPases in the development of cell polarity in yeast and beyond.' Microbiol Mol Biol Rev 71(1): 48-96. [21] Sanders, S. L. and I. Herskowitz (1996). 'The BUD4 protein of yeast, required for axial budding, is localized to the mother/BUD neck in a cell cycle-dependent manner.' J Cell Biol 134(2): 413-427. [22] Si, H., D. Justa-Schuch, et al. (2010). 'Regulation of septum formation by the Bud3-Rho4 GTPase module in Aspergillus nidulans.' Genetics 185(1): 165-176. [23] Stegmeier, F. and A. Amon (2004). 'Closing mitosis: the functions of the Cdc14 phosphatase and its regulation.' Annu Rev Genet 38: 203-232. [24] Taylor, G. S., Y. Liu, et al. (1997). 'The activity of Cdc14p, an oligomeric dual specificity protein phosphatase from Saccharomyces cerevisiae, is required for cell cycle progression.' J Biol Chem 272(38): 24054-24063. [25] Ubersax, J. A., E. L. Woodbury, et al. (2003). 'Targets of the cyclin-dependent kinase Cdk1.' Nature 425(6960): 859-864. [26] Visintin, R., K. Craig, et al. (1998). 'The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk-dependent phosphorylation.' Mol Cell 2(6): 709-718. [27] Zachariae, W. and K. Nasmyth (1999). 'Whose end is destruction: cell division and the anaphase-promoting complex.' Genes Dev 13(16): 2039-2058. [28] Zachariae, W., M. Schwab, et al. (1998). 'Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex.' Science 282(5394): 1721-1724. [29] Zheng, Y., A. Bender, et al. (1995). 'Interactions among proteins involved in bud-site selection and bud-site assembly in Saccharomyces cerevisiae.' J Biol Chem 270(2): 626-630. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46436 | - |
| dc.description.abstract | 蛋白後修飾在生物細胞功能中扮演重要角色，後修飾包含有磷酸化、甲基化、乙醯化…等等，其中，磷酸化為目前研究最為透徹的後修飾作用。在酵母菌細胞中，Cdk1是最有力的激酶，在細胞週期中，Cdk1的活化需與不同的cyclin相互配合形成復合物，進而可磷酸化不同受質、甚至是受質的調控蛋白，進而影響整個酵母菌細胞週期，許多Cdk1的受質已經被報導出來，像是Sic1、Cln2、
Swi5…等。另外，要脫離一個細胞週期，則需要Cdc14酵素去磷酸化的作用，細胞走到M時期時，細胞染色體合成結束、分離後，就準備要脫離、進入下一個細胞週期，Cdc14從核仁釋出、移動至目標蛋白來進行去磷酸化的功能，像是與Sic1作用，因而可以減低Cdk1活性，使細胞準備好要脫離有絲分裂期。不同於Cdk1，很少的Cdc14受質被報導，因此我們有興趣知道是否有任何受質同時受到Cdk1及Cdc14兩種酵素的調控，進而影響到酵母菌的表現型。運用反向思考，我們分別收集野生型、溫度感知的Cdc14缺陷型酵母菌蛋白，並做分層處理，隨後針對磷酸化層的蛋白做質譜分析。從質譜分析得來的結果，相較於野生型，挑選出具有較高磷酸態的蛋白，進一步則是將之對比於Cdk1磷酸化的consensus sties，從其中篩選出一些候選蛋白。而BUD3是其中一個蛋白，我們將研究它是否真是同時受Cdk1及Cdc14調控的受質，並探討這樣的調控對酵母菌有任何功能影響。 | zh_TW |
| dc.description.abstract | Cdc28 is well-known as the catalytic subunit of main cyclin- dependent kinase, whose activity associates with different substrates would drive events through cell cycle. Many substrates of Cdc28 have been reported, such as Sic1, Cln2, and Swi5…, etc. The protein phosphatase, Cdc14, required for mitotic exits. With anaphase onset, Cdc14 released from nucleolus by FEAR network and mitotic exit network, which enable a decrease in Cdk-Clb activity. Therefore, we wondered if there any unknown substrates of Cdc28 would be found. We had compared the phosphorylation level of proteins between wild type and cdc14 defective mutant strain by mass-spectrometry. After scoring, we got some candidates which have higher phosphorylation level in cdc14 mutant than wild type. The following steps are going to identify and characterize those candidates. BUD3 is one of the candidates from the Mass-data which is expressed close to the onset of mitosis and required for the axial pattern of cell division. It has shown that there are two sites located on the C-terminal possibly regulated by Cdc28 and Cdc14. We want to examine whether phosphorylation level of Bud3 control by Cdc28 and Cdc14. Then, we’ll going to perform both in vivo and in vitro assays to verify the relationship between Bud3, Cdc28 and Cdc14, and what events they’ll contribute to in the cell cycle of a budding yeast. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T05:08:56Z (GMT). No. of bitstreams: 1 ntu-99-R97445107-1.pdf: 1862339 bytes, checksum: ebeb76b8bdcf507fe88b102f0ce890df (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | CONTENTS
口試委員會審定書 # 誌謝 1 中文摘要 2 ABSTRACT 3 CONTENTS 4 Chapter 1 Introduction 6 Chapter 2 Material and methods 9 2.1 Yeast strains 9 2.2 Cell extracts and western blotting analysis 10 2.3 Design and generation of phospho-specific antibodies 11 2.4 In vitro Immuno-precipitation kinase assay 11 2.5 Recombinant protein purification 12 2.6 Staining of Bud scars by Calcofluor 12 Chapter 3 Results 14 3.1 Analysis of LC-Mass/Mass 14 3.2 The residues in C-terminal of BUD3 are phosphorylated by CDK1 in vitro 15 3.3 Generation of phosphor-specific antibodies and detection of endogenous BUD3 15 3.4 Both residues, S1549 and T1566, might be the substrates of Cdc14 and Cdk1 16 3.5 Mutations of the residues S1549, T1566, and S1549/T1566 might have effect on budding pattern of the budding yeast 17 3.6 Mutation of the residues S1549, T1566, and S1549/T1566 will not affect the interaction with CLB2; the autoregulation between C-terminal and GEF of BUD3 was not found 18 Chapter 4 Discussion 20 Chapter 5 Figures 23 5.1 Figure.1(a) LC-MS/MS analysis 23 5.2 Figure.1(b) LC-MS/MS analysis 24 5.3 Figure.2 Both BUD3 residues, S1549 and T1566, are phosphorylated by CDK1 in vitro 25 5.4 Figure.3 Generation of phosphor-specific antibodies 26 5.5 Figure.4 Both BUD3 residues, S1549 and T156, are dephosphorylated by CDC14 in vivo 27 5.6 Figure.5 The residue of BUD3, S1549, is phosphorylated by CDK1 in vivo 28 5.7 Figure.6 Calcoflour staining observed by excitatory illumination 29 5.8 Figure.7 Quantitative analysis of bipolar pattern 30 5.9 Figure.8 BUD3 C-terminal did not associate with CLB2 in yeast-two hybrid system 31 5.10 Figure.9 There was no association between the C-terminal and GEF domain of BUD3 32 TABLES 33 REFERENCE 34 | |
| dc.language.iso | en | |
| dc.subject | 蛋白後修飾 | zh_TW |
| dc.subject | 第二出芽位置選擇 | zh_TW |
| dc.subject | Bud3 | en |
| dc.subject | Cdk1 | en |
| dc.subject | Cdc14 | en |
| dc.title | 在Saccharomyces cerevisiae酵母菌模式中,Bud3是否經由Cdk1調控而影響第二出芽位置選擇的方向 | zh_TW |
| dc.title | Cdk1-mediated Bud3 regulation of bud site selection in Saccharomyces cerevisiae | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李財坤(Tsai-Kun Li),詹迺立(Nei-Li Chan) | |
| dc.subject.keyword | 第二出芽位置選擇,蛋白後修飾, | zh_TW |
| dc.subject.keyword | Cdk1,Cdc14,Bud3, | en |
| dc.relation.page | 36 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-07-26 | |
| 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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