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博碩士論文 etd-0827108-124120 詳細資訊
Title page for etd-0827108-124120
論文名稱
Title
Sp1的磷酸化增強與HDAC1結合而抑制RECK表現
Phosphorylation of Sp1 enhances HDAC1 recruitment to inhibit RECK expression
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
60
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2008-07-23
繳交日期
Date of Submission
2008-08-27
關鍵字
Keywords
Sp1、HDAC1、RECK
磷酸化
統計
Statistics
本論文已被瀏覽 5689 次,被下載 12
The thesis/dissertation has been browsed 5689 times, has been downloaded 12 times.
中文摘要
轉錄因子Sp1可以藉由和不同的共同轉錄活化或抑制因子的結合進而對多種基因表現做正向和負向調控。在我們先前的實驗顯示Her2/neu致癌基因會經由ERK/Sp1 訊息活化路徑抑制腫瘤抑制基因RECK (reversion-inducing cysteine rich protein with Kazal motifs) 的表現,而且當HDAC1抑制物trichostatin A的作用下可回復RECK的表現及減少癌症細胞的轉移。因此,我們的目的想探討Sp1 和HDAC1的結合與RECK基因轉錄的機制。首先我們藉由GST pull-down assay發現Sp1上的zinc-finger DNA binding domain是和HDAC1結合的重要區域。再進一步以co-immunoprecipitation、luciferase assay和RT-PCR實驗顯示Sp1在Thr739 區位磷酸化具有可以影響和HDAC1的結合進而調控RECK的轉錄表現能力。基於以上結果我們推測藉由ERK/Sp1訊息路徑磷酸化Sp1上的Thr739區位可以吸引HDAC1的結合來抑制RECK轉錄表現。
Abstract
Transcription factor Sp1 regulates both negatively and positively the expression of target genes by interacting different co-activator or co-repressor. Our previous studies showed that Her2/neu oncogene inhibited RECK (reversion-inducing cysteine rich protein with Kazal motifs)transcription via the ERK/Sp1 signaling pathway and histone deacetylase (HDAC) inhibitor, trichostatin A(TSA), reversed expression of RECK and reduced cancer cell metastasis. Therefore, we purposed that RECK gene transcription activity was controlled by Sp1 and HDAC1 interaction. By GST pull-down assay, we discovered that zinc-finger DNA binding domain is a crucial site which affected Sp1 binding with HDAC1. Further investigation revealed that the phosphorylation at Thr739 residue of Sp1 regulated the level of RECK transcription as determined by co-immunoprecipitation、luciferase assay and RT-PCR. Based on these results, we suggest that phosphorylation at Thr739 of Sp1 by the ERK/Sp1 signaling pathway can recruit HDAC1 to inhibit RECK gene trancription.
目次 Table of Contents
中文摘要 2
英文摘要 3
縮寫語 4
前言 5
實驗材料與方法 9
實驗結果 23
討論 28
實驗結果圖表 33
參考文獻 43
附圖 52
參考文獻 References
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