2016-01-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657922摘要：陰道滴蟲引起之陰道滴蟲症是最常見非病毒源性病，感染可能促進人類免疫不全病毒與 乳突病毒傳播率，突顯其公衛學上重要性。先期研究證實鐵離子引發訊息傳導，快速調節Myb3 轉錄因子入核；同時，發現鐵可能誘導組織蛋白去乙醯酶(Histone deacetylase 1, TVHDAC1)之 Ser391磷酸化上升，相反地組織蛋白Histone之H3K9乙醯化下降，兩者皆被認為與人類表徵 遺傳調控抑制基因轉錄相關；推測鐵引發陰道滴蟲訊息傳導活化TVHDAC1，可能導致組織蛋 白去乙醯化，進一步調控表徵遺傳之基因轉錄。為了證實此一假說，計晝以蛋白質體學策略 分析陰道滴蟲受鐵誘發後全細胞蛋白質的磷酸化變化。此外，從酵素活性、蛋白質表現、細 胞核質分佈等現象，探討TVHDAC1基礎生化特性，並以定量蛋白質體學分析確認TVHDAC1 功能，在鐵刺激組織蛋白各式轉譯後修飾變化中扮演的角色。同時以高通量定序染色質體免 疫沉澱產物，分析細胞受鐵刺激後TVHDAC1對DNA結合模式，以分析結合區域之基因分佈， 並以即時定量反轉錄聚合酶鏈鎖反應確認此些基因表現是否真實受鐵調控。本計晝所提出全 新的概念，為陰道滴蟲鐵誘發表徵遺傳調控研究提供重要訊息。<br> Abstract: Human pathogen, Trichomonas vaginalis is an etiologic agent of trichomoniasis, the most prevalent non-viral sexually transmitted disease, which has been linked to elevated transmission of the human immunodeficiency virus and papillomaviruses. In previous reports, iron was shown to rapidly trigger signal transduction and accelerate nuclear import of a Myb3 transcription factor. In the preliminary study, iron was found to increase phosphorylation of histone deacetylase 1TQ1(TvHDAC1) at Ser and deacetylation of histone H3K9. Both types of modifications are known as epigenetic hallmarks of gene silencing in human, suggesting that iron may sequentially promotes TvHDAC1 activation, histone H3K9 deacetylation and epigenetic interference of gene expression. In this proposal, a proteomics approach shall be conducted in triplicate to identify global phosphorylation profiles of cellular proteins in response to iron overloading. In addition, biochemical characterization of TvHDAC 1 like its enzyme activity in vitro and its responses to iron overloading shall be examined exploring a transgenic protein expression system. Moreover, the role of TvHDAC1 in iron-inducible histone modifications shall be examined by a proteomics approach. Ultimately, genome-wide targeting-sites of TvHDAC1 shall be mapped by a chromatin-immunoprecipitation based approach. Iron-inducible or -suppressive gene expression shall be examined by the quantitative reverse transcriptase-polymerase chain reaction assay. Since little is known on the regulation of gene expression in T. vaginalis, the study proposed herein shall provide sound ground for our understandings on epigenetic regulation in the parasite.陰道滴蟲組織蛋白去乙醯酶組織蛋白質鐵離子表徵遺傳調控訊息傳導Trichomonas vaginalisHistone deacetylaseHistoneIronEpigenetic regulationSignal transduction