English  |  正體中文  |  简体中文  |  全文筆數/總筆數 : 45422/58598 (78%)
造訪人次 : 2528931      線上人數 : 187
RC Version 7.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
搜尋範圍 查詢小技巧：
  • 您可在西文檢索詞彙前後加上"雙引號"，以獲取較精準的檢索結果
  • 若欲以作者姓名搜尋，建議至進階搜尋限定作者欄位，可獲得較完整資料
    •  進階搜尋


    請使用永久網址來引用或連結此文件: http://libir.tmu.edu.tw/handle/987654321/5892


    題名: 探討活化型態的Notch1受體與其結合因子之結合生物功能
    Study on the biological function of the interaction between activated Notch1 receptor and its binding factors
    作者: 林育民
    Yu-Min Lin
    貢獻者: 細胞及分子生物研究所
    關鍵詞: Notch1受體
    Notch1
    日期: 2003
    上傳時間: 2009-09-11 15:46:34 (UTC+8)
    摘要: 中文摘要
    Notch是個演化上高度保留，穿過細胞膜一次的受體蛋白，最早在果蠅中被發現。已知哺乳類的Notch受體蛋白分為Notch1?4，位於細胞表面，被認為可調控基因的轉錄、細胞的增生、分化及早期細胞命運的決定，而Notch受體蛋白異常的活化會造成某些疾病的發生。當相鄰細胞表現的ligand結合在Notch受體蛋白的細胞外區域時，Notch訊號傳遞系統就被活化，使得整個細胞內區域在靠近內膜處斷裂，而形成活化型Notch (Notch IC, 細胞內區域)。此活化型Notch會轉移至細胞核中，並且和一些可與DNA結合的轉錄因子，如：RBP-Jκ/CBF1，以及其他細胞因子結合，進一步調控其標的基因的表現。由於這些參與的因子，對於Notch訊息傳遞路徑影響甚巨。因此，為了進一步探討Notch訊息傳遞路徑，本論文首要目標著重於探索參與Notch1下游訊息傳遞路徑仍未被發現的結合因子，及初步分析因結合所產生的生物功能。
    在實驗的第一部分，我們利用K562、Jurkat、SUP-T1等人類細胞株證實了一個轉錄因子YY1與Notch1 IC (N1IC)之間的結合關係，同時也利用GST pull-down assay，找出N1IC和YY1 相互之間的結合區域，並藉由Sucrose gradient analysis證實細胞核內N1IC及YY1以大的complex結合形式存在，本論文也證實 YY1藉由N1IC與CBF1的直接作用結合在Wild type CBF1-response elements上，YY1會抑制CBF1媒介之Notch1訊息傳遞。同時，應用ChIP嘗試尋找未知受人類Notch1訊息傳遞所調控的下游基因。綜觀上述實驗結果，我們得知YY1藉由與N1IC結合，形成高分子量的複合體調控Notch 1訊息傳遞。
    在實驗的第二部分，延續之前本實驗室已證實N1IC與b2-tubulin之間相互結合的結果，應用上述細胞分析N1IC與b2-tubulin之間的結合生物功能。實驗結果證實N1IC與b2-tubulin之間的結合是同時發生於細胞核及細胞質的，而應用RNA interference分析及細胞處理taxol的實驗結果更進一步證實b2-tubulin可能抑制經CBF1媒介之Notch1訊息傳遞。
    Abstract
    Notch receptors are evolutionally highly conserved and play important roles in modulating cell fate decisions throughout the development from invertebrate to vertebrate species. Four homologues of Notch receptors have been identified in human and aberrant Notch signaling is associated with a number of human diseases. There is a consensus model of Notch signaling pathway containing multiple proteolytic steps. After ligand binding, Notch receptors are cleaved to release the intracellular domains of Notch receptors. The intracellular domains, the activated form of Notch receptors, are then translocated into nuclei and interact with other transcriptional machineries to regulate the expression of cellular genes.
    To dissect the molecular mechanisms of Notch signaling, the cellular targets interact with Notch 1 receptor intracellular domain (N1IC) were screened. In this study, we found that endogenous transcription factor Ying Yang 1 (YY1) was associated with exogenous N1IC in human K562 erythroleukemic cells. The ankyrin domain of N1IC and zinc finger domains of YY1 were essential for the association of N1IC and YY1 by the pull-down assay of GST fusion proteins. Furthermore, both YY1 and N1IC were present in a large complex of the nucleus to suppress the luciferase reporter activity trans-activated by Notch signaling. The transcription factor YY1 indirectly regulated the transcriptional activity of the wild-type CBF1-response elements via the direct interaction of N1IC and CBF1. We also demonstrated the association between endogenous N1IC and intrinsic YY1 in human Jurkat cells and T-cell acute lymphoblastic leukemia cells. Additionally, we identified the cellular target genes modulated by the high-molecular-weight Notch complex by chromatin immunoprecipitation. Taken together, these results indicate transcription factor YY1 may modulate Notch signaling via association with the high-molecular-weight Notch complex.
    We also demonstrated that the microtubule component 2-tubulin was associated with exogenous and endogenous N1IC in cells, Further study showed that N1IC and 2-tubulin associated in both nucleus and cytosol. The expression of 2-tubulin was inhibited by RNA interference and the nuclear 2-tubulin was depleted by the treatment of taxol to investigate the roles of 2-tubulin in Notch1 signaling pathway. The results suggested that 2-tubulin suppresses the luciferase reporter activity trans-activated by Notch1 signaling.
    資料類型: thesis
    顯示於類別:[醫學科學研究所] 博碩士論文

    文件中的檔案:

    檔案 描述 大小格式瀏覽次數
    摘要.doc30KbMicrosoft Word94檢視/開啟
    摘要.pdf68KbAdobe PDF65檢視/開啟
    摘要.ppt109KbMicrosoft Powerpoint49檢視/開啟
    摘要.ps410KbPostscript56檢視/開啟


    在TMUIR中所有的資料項目都受到原著作權保護.

    TAIR相關文章

    著作權聲明 Copyright Notice

    • 本平台之數位內容為臺北醫學大學所收錄之機構典藏，包含體系內各式學術著作及學術產出。秉持開放取用的精神，提供使用者進行資料檢索、下載與取用，惟仍請適度、合理地於合法範圍內使用本平台之內容，以尊重著作權人之權益。商業上之利用，請先取得著作權人之授權。

      The digital content on this platform is part of the Taipei Medical University Institutional Repository, featuring various academic works and outputs from the institution. It offers free access to academic research and public education for non-commercial use. Please use the content appropriately and within legal boundaries to respect copyright owners' rights. For commercial use, please obtain prior authorization from the copyright owner.

    • 瀏覽或使用本平台，視同使用者已完全接受並瞭解聲明中所有規範、中華民國相關法規、一切國際網路規定及使用慣例，並不得為任何不法目的使用TMUIR。

      By utilising the platform, users are deemed to have fully accepted and understood all the regulations set out in the statement, relevant laws of the Republic of China, all international internet regulations, and usage conventions. Furthermore, users must not use TMUIR for any illegal purposes.

    • 本平台盡力防止侵害著作權人之權益。若發現本平台之數位內容有侵害著作權人權益情事者，煩請權利人通知本平台維護人員([email protected])，將立即採取移除該數位著作等補救措施。

      TMUIR is made to protect the interests of copyright owners. If you believe that any material on the website infringes copyright, please contact our staff([email protected]). We will remove the work from the repository.

    Back to Top
    DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - 回饋