2008-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/685868摘要：EB病毒表現的溶裂期極早期Rta蛋白質會活化溶裂期基因的表現及溶裂循環。最近利用酵母菌雙雜交分析發現，一種Ran nuclear-cytoplasmic transport protein, RanBPM 會與Rta結合。藉由GST pull-down assay 也證明RanBPM與Rta會結合。然後用共免疫沈澱法及免疫螢光分析發現RanBPM與Rta在細胞內會結合並共同位在細胞核內。另外，利用細胞轉染分析證明RanBPM與Rta的結合會活化病毒BMLF1及細胞內p21基因的轉錄作用。RanBPM也會與SUMO的E2 (Ubc9)結合。另外我也發現RanBPM會與EB病毒另ㄧ個重要的極早期蛋白質Zta在體外直接結合，顯示RanBPM很可能會影響Zta與Rta對EB病毒溶裂循環的活化。基於目前這些研究結果，本研究計畫的目的將要證明(1) RanBPM對Rta sumoylation的活化機制；(2) RanBPM與Zta的結合作用；及(3) RanBPM與Zta的結合如何影響EB病毒溶裂循環的進行。這個研究將會更清楚瞭解病毒與宿主的蛋白質是如何交互作用而使病毒能夠增殖。<br> Abstract: Epstein-Barr virus (EBV) expresses a transcription factor, Rta, during the immediate-early stage of the lytic cycle to activate the transcription of the EBV lytic genes and the lytic cycle. My recent yeast two-hybrid analysis showed that RanBPM, a Ran nuclear-cytoplasmic transport protein, is a binding partner of Rta. The binding was confirmed by glutathione S-transferase pull-down assay. A coimmunoprecipitation experiment and confocal microscopy study revealed that RanBPM and Rta interact in vivo and colocalize in the nucleus. The interaction promotes the transactivation activity of Rta in activating the transcription of BMLF1 and p21 in transient transfection assay. Additionally, RanBPM interacts with SUMO-E2 (Ubc9). Moreover, my study showed that Zta, another immediate-early protein of EBV, interacts with RanBPM in vitro, implying that RanBPM affects the capacity of both Zta and Rta to activate the EBV lytic cycle. Based on these findings, the studies proposed herein will investigate (1) the mechanisms by which RanBPM enhances Rta sumoylation; (2) the interaction between Zta and RanBPM; and (3) how the interaction between RanBPM and Zta influences EBV lytic development. This work will lead to a better understanding on how cellular and viral proteins cooperate to enable the virus to proliferate.RanBPMEpstein-Barr virusRtaZtalytic cycle