靳宗洛臺灣大學:植物科學研究所賴慧娟Lai, Hui-ChuanHui-ChuanLai2010-05-112018-07-062010-05-112018-07-062009U0001-1507200914474900http://ntur.lib.ntu.edu.tw//handle/246246/181971熱休克反應，為生物體內普遍存在的機制；藉由誘導熱休克蛋白質的生成，進行細胞保護，而熱休克蛋白質的生成是經由熱休克轉錄因子所調控。人類熱休克轉錄因子結合蛋白(HsHSBP1)可以和熱休克轉錄因子進行交互作用，終止熱休克反應，扮演負向調控者的角色。關於植物HSBP參與熱休克反應的機制目前並不清楚。藉由序列分析，我們在阿拉伯芥中篩選到一個HSBP基因(AtHSBP)，並分離出兩個基因缺陷突變株。經由測試其熱耐受性，顯示突變株具有較高的「誘導耐熱性」，但是對於「先天耐熱性」並沒有顯著的影響。在阿拉伯芥原生質體中表現GFP-AtHSBP，得知在正常生長情況以及熱處理的初期，AtHSBP表現於細胞質中；但長時間的熱處理及當其回復到正常生長情況時，AtHSBP會自細胞質轉移至細胞核。雙分子螢光互補(Bimolecular fluorescence complementation)分析的結果顯示，AtHSBP會和AtHSFA1a, AtHSFA1b以及AtHSFA2進行交互作用。利用即時定量聚合酶鍊鎖反應(quantitative real-time PCR)分析，得知AtHSBP缺陷突變株在遭受熱逆境時，熱休克蛋白基因(HSP101, HSP70, HSP17.4, HSP18.2, and HSP25.3)的表現量及累積量比野生型多；而AtHSBP的過度表現株，其熱休克蛋白基因的表現量及累積量下降，並導致其「誘導耐熱性」能力降低。經由以上結果，我們推測AtHSBP在熱休克反應對於熱休克蛋白的表現，扮演負向調控者的角色。Abstract in Chinese---------------------------------------------------------------------------- 1bstract in English---------------------------------------------------------------------------- 2bbreviations----------------------------------------------------------------------------------- 3ntroduction------------------------------------------------------------------------------------- 4eat shock response and heat shock proteins----------------------------------------------- 4egulation of heat shock response ----------------------------------------------------------- 5eat shock factors in Arabidopsis------------------------------------------------------------ 6revious study of heat shock factor binding proteins-------------------------------------- 8oals and strategies of the dissertation------------------------------------------------------ 9aterials and Methods---------------------------------------------------------------------- 11lant material, transformation and growth condition ------------------------------------- 11NA extraction and quantitative real-time PCR ------------------------------------------ 11hermotolerance phenotypes assay --------------------------------------------------------- 12ransient expression in Arabidopsis protoplast-------------------------------------------- 12ubcellular localization and bimolecular fluorescence complementation assay ------13esults------------------------------------------------------------------------------------------ 14equence alignment and phylogenetic analysis of the HSBP homologs across plants and animals------------------------------------------------------------------------------------- 14xpression profiles of AtHSBP in various tissues and with HS treatment ------------ 15hermotolerance analysis in AtHSBP mutants-------------------------------------------- 15tHSBP is a cytosolic-localized protein capable of entering nucleus in response to thermal stress----------------------------------------------------------------------------------- 16tHSBP interacts with AtHSFA1a, AtHSFA1b and AtHSFA2 in vivo----------------- 17he HSP expression patterns in AtHSBP mutants during HSR------------------------- 18iscussion-------------------------------------------------------------------------------------- 19tHSBP is a conserved protein in HSBP family and may participate in embryogenesis and environmental stress---------------------------------------------------------------------- 19tHSBP acts as a negative regulator during HSR----------------------------------------- 20tHSBP participates in the temporal and spatial regulation of HSP genes expression under the influence of heat stress------------------------------------------------------------ 22igures and Table----------------------------------------------------------------------------- 23eferences-------------------------------------------------------------------------------------- 35upplemental data---------------------------------------------------------------------------- 40application/pdf1068496 bytesapplication/pdfen-US熱休克轉錄因子結合蛋白熱逆境熱休克轉錄因子阿拉伯芥HSBPheat stressHSFArabidopsishttp://ntur.lib.ntu.edu.tw/bitstream/246246/181971/1/ntu-98-R95b42003-1.pdf