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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張智芬 | |
| dc.contributor.author | Yuan-Yeh Kuo | en |
| dc.contributor.author | 郭遠燁 | zh_TW |
| dc.date.accessioned | 2021-06-13T01:36:29Z | - |
| dc.date.available | 2007-07-20 | |
| dc.date.copyright | 2007-07-20 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-15 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30104 | - |
| dc.description.abstract | Gfi-1B屬於Gfi家族之轉錄抑制因子(transcriptional repressor），是紅血球新生作用(erythropoiesis)過程的必要分子。在紅血球前驅細胞 (erythroid progenitor cells)中，表現大量Gfi-1B，可以造成這些細胞在7至10天後大量凋亡(apoptosis) 的現象。同時，抗凋亡蛋白質Bcl-xL的表現量也明顯的減少。因此，本研究首要目的，在於瞭解Gfi-1B在Bcl-xL之轉錄調節之角色。
由於GATA-1亦是一個紅血球生成之必要轉錄因子，本實驗室過去的研究發現Gfi-1B可和GATA-1結合。因此，在本論文研究中，利用染色質沈澱分析法(ChIP)得知，在紅血球前驅細胞分化過程中，GATA-1始終都與Bcl-x的啟動子(promoter)結合，Gfi-1B只有分化前期才會與Bcl-x的啟動子結合。GATA-1可結合在Bcl-x啟動子上的GATT位置，誘發Bcl-xL基因表現，過度表現Gfi-1B則可以抑制GATA-1對Bcl-xL的活化。Gfi-1B是透過與GATA-1的結合而被帶領到Bcl-x啟動子上，並進行轉錄抑制。此外，在K562細胞加入Bcr-Abl激酶抑制劑Imatinib可以導致Gfi-1B的表現量上升。此時，Gfi-1B也可與GATA-1結合並抑制Bcl-xL的基因表現。利用干擾性核醣核酸(RNA interference)降低Gfi-1B的表現可以減少Imatinib所引發的細胞凋亡。另一方面，在K562細胞中過度表現Gfi-1B則會使細胞對砷化物(arsenic)所引發的死亡更為敏感。以上的實驗結果，闡釋了Gfi-1B在紅血球細胞內協同GATA-1參與誘導細胞凋亡之角色。 此外，本論文亦研究Gfi-1B的後轉錄調控機制(post-transcriptional control)，並發現Gfi-1B的訊息核醣核酸(mRNA)半衰期短，該Gfi-1B mRNA具一可造成該核醣核酸不穩定的片段。同時，其 mRNA上之負面調節因子是有賴於轉譯作用以利RNA分解之進行。 | zh_TW |
| dc.description.abstract | Gfi-1B (growth factor independence-1B) gene is a Gfi-family transcriptional repressor, whose expression plays an essential role in erythropoiesis. Early erythroid progenitor cells overexpressing Gfi-1Bexhibit massive apoptosis after 7-10 days of culture with a significant reduction of Bcl-xL expression. Therefore, the aim of this study is to uncover the role of Gfi-1B in the regulation of Bcl-xL transcription
GATA-1 is also an essential transcriptional factor in erythropoiesis. Our laboratory previously showed that Gfi-1B interacts with GATA-1. Using ChIP assays, I provide evidence that GATA-1 binds to the Bcl-x promoter constantly in the entire induction period, while Gfi-1B is transiently associated with the promoter in the early phase. GATA-1 binds to the noncanonical GATT motif of the Bcl-x promoter for trans-activation, and that enforced expression of Gfi-1B represses the Bcl-x promoter in a GATA-1-dependent manner. Gfi-1B expressed at increased levels is recruited to the Bcl-x promoter through its association with GATA-1, suppressing Bcl-xL transcription. Furthermore, I showed that imatonob, Bcr-Abl kinase inhibitor, causes up-regulation of Gfi-1B in K562 cells, where it also cooperated with GATA-1 to repress Bcl-xL transcription. Gfi-1B knockdown by RNA interference diminished imatinib-induced apoptosis, while the overexpression of Gfi-1B sensitized K562 cells to arsenic-induced death. These findings illuminate the role of Gfi-1B in GATA-1-mediated transcription in the survival aspect of erythroid cells. In addition, the post-transcriptional control of the Gfi-1B gene is studied. It was found that Gfi-1B mRNA is a short half-life transcript containing a destabilizing element within the coding region of the Gfi-1B mRNA. I further defined the region of the Gfi-1B coding region determinant (CRD) and demonstrated that this CRD confers cis-directed destabilization coupled with translation. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T01:36:29Z (GMT). No. of bitstreams: 1 ntu-96-D90442001-1.pdf: 2841597 bytes, checksum: 86b182b42f9df99e96816bacbdff1555 (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 論文口試委員審定書……………………………………………………i
謝誌…………………………………………………………………… ii 中文摘要………………………………………………………………iii Abstract ……………………………………………………………… iv Preface ……………………………………………………………… 1 Chapter I- Overview and Rationale Overview of erythropoiesis……………………………………… 2 Role of GATA-1 in normal hematopoiesis……………………… 3 Gfi-1 family ………………………………………………………… 6 Overview of mRNA degradation…………………………………… 10 Experimental Rationale …………………………………………14 Chapter II- GATA-1 and Gfi-1B Interplay to Regulate Bcl-xL Transcription Introduction …………………………………………………………16 Materials and Methods ………………………………………………19 Results …………………………………………………………………24 Discussion ……………………………………………………………33 Figures & Legends……………………………………………………37 Chapter III-A Coding Region Determinant Regulates stability of Gfi-1B mRNA Introduction ………………………………………………………52 Materials and Methods ……………………………………………56 Results ………………………………………………………………60 Discussion …………………………………………………………65 Figures & Legends…………………………………………………67 References……………………………………………………………75 | |
| dc.language.iso | en | |
| dc.subject | 核酸穩定性 | zh_TW |
| dc.subject | 轉錄抑制 | zh_TW |
| dc.subject | transcriptional repression | en |
| dc.subject | RNA stability | en |
| dc.title | Gfi-1B於造血細胞內之轉錄抑制功能及其核酸穩定性之調節機制 | zh_TW |
| dc.title | The Transcriptional Repression Function and RNA Stability Control of Growth Factor Independence-1B (Gfi-1B) in Hematopoietic Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 嚴仲陽,阮麗蓉,繆希椿,羅?升 | |
| dc.subject.keyword | 轉錄抑制,核酸穩定性, | zh_TW |
| dc.subject.keyword | transcriptional repression,RNA stability, | en |
| dc.relation.page | 84 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-07-16 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
| 顯示於系所單位： | 生物化學暨分子生物學科研究所 | |
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