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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄧述諄(Shu-Chun Teng) | |
dc.contributor.author | Yu-Ching Huang | en |
dc.contributor.author | 黃裕清 | zh_TW |
dc.date.accessioned | 2021-06-13T02:03:38Z | - |
dc.date.available | 2021-08-01 | |
dc.date.copyright | 2011-10-07 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-02 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30430 | - |
dc.description.abstract | 細胞週期中的G1到S期的轉換大部份是靠G1週期素和Cdc28週期素依賴性蛋白激酶複合體的活性來調控。G1週期素中的Cln1和Cln2對於細胞週期的啟動提供了特殊的貢獻,而Cln3則是作用在CLN1和CLN2時間點上的表現和其他在G1時期表現的基因。之前的研究顯示出Ypl014w是G1週期素/Cdc28複合體的其中一員。然而,這個基因是如何在細胞週期中被調控以及是否影響G1週期素–週期素依賴性蛋白激酶複合體的活性仍然是未知的。在本篇論文中,我們發現YPL014W的表現模式是呈現細胞週期性地調控。我們顯示YPL014W是在細胞週期中的G1時期表現,並且它的表現是靠轉錄因子Mcm1的直接接合。在YPL014W的啟動子內有兩個賦予M/G1時期基因轉錄的early cell cycle box (ECB)位置,而這兩個位置對於YPL014W週期性地表現是很重要的。當Ypl014w大量表現時,細胞週期大規模地停滯在G1時期。除此之外,我們用共同免疫沉澱法證實了Ypl014w和G1週期素是會互相結合的。G1和S週期素依賴性蛋白質激酶的活性可直接被Ypl014w抑制。我們同時也發現,當粒腺體失去功能時將會促進YPL014W基因的表現量上升。本篇論文證明了Ypl014w可能是週期素依賴性蛋白激酶的抑制物,而且是在細胞遭受粒腺體壓力時作用在調控細胞週期的進行。 | zh_TW |
dc.description.abstract | The G1-S transition in the cell cycle is largely regulated by the activity of the G1 cyclin-Cdc28 cyclin-dependent protein kinase complex. Among G1 cyclins, Cln1 and Cln2 appear to make a specific contribution to the initiation of the cell cycle, while Cln3 functions for the timely expression of CLN1, CLN2 and other G1-specific genes. Previous results suggest that Ypl014w is a component of the G1 cyclin/Cdc28 complex. However, it is still elusive how this gene is cell cycle-regulated and whether it affects the activity of the G1 cyclin-CDK complex. Here we show that YPL014W is expressed in a cell cycle-regulated pattern. We found that YPL014W is expressed in G1 phase of the cell cycle and is activated by direct binding of the transcription factor, Mcm1. Two early cell cycle box (ECB) elements, which confer M/G1-specific transcription, were necessary for the periodicity expression of YPL014W. When Ypl014w was overexpressed, cell cycle ceased dominantly at G1 stage. In addition, the interactions between Ypl014w and G1 cyclins were confirmed by co-immunoprecipitation. The kinase activity of G1 cyclins and S phase cyclins/CDK was directly inhibited by Ypl014w. Mitochondrial dysfunction induces up-regulation of YPL014W, thus demonstrating that Ypl014w may act as a CDK inhibitor to modulate cell cycle progression when cells encounter mitochondrial stress. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T02:03:38Z (GMT). No. of bitstreams: 1 ntu-100-F94445114-1.pdf: 2396605 bytes, checksum: 24fb3f74135bcb92e42809a34de58fd6 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | INTRODUCTION----------1
MATERIALS & METHODS----------7 Yeast strains and plasmids----------7 Cell synchronization and protein analysis----------8 RNA analysis----------8 Chromatin immunoprecipitation (ChIP) analysis----------9 Reverse transcription and real-time PCR assay----------9 Protein sequence alignment and secondary structure prediction----------10 Recombinant protein purification----------10 Kinase assay----------10 RESULTS----------12 SIC2 is expressed in G1 stage during the cell cycle----------12 Mcm1 regulates SIC2 expression before the G1 stage----------12 Mcm1 directly associates with the two ECB elements at the promoter of SIC2----------13 Overexpression of SIC2 induces G1 arrest----------15 The middle domain of Sic2, which contains two alpha-helices, is highly conserved in yeast species----------17 The two alpha-helices are both critical for Sic2-mediated growth retardation----------18 Association of Sic2 with Cln1, Cln2, and Cln3----------18 Sic2 inhibits G1 cyclin/CDK protein kinase activity in vitro----------19 Mitochondrial stress induces SIC2 expression----------20 DISCUSSION----------23 FIGURES----------26 REFERENCES----------40 APPENDIX----------49 | |
dc.language.iso | en | |
dc.title | Ypl014w在出芽酵母菌中為一新穎的細胞週期磷酸酶抑制物 | zh_TW |
dc.title | Ypl014w, a novel CDK inhibitor of the cell cycle in budding yeast | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 李財坤(Tsai-Kun Li),詹世鵬(Shih-Peng Chan),詹迺立(Nei-Li Chan),張壯榮(Chuang-Rung Chang) | |
dc.subject.keyword | 細胞週期,磷酸酶,抑制物,粒腺體, | zh_TW |
dc.subject.keyword | cell cycle,CDK inhibitor,mitochondria, | en |
dc.relation.page | 49 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-02 | |
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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