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Title: 以斑馬魚為模式動物研究魚類精胺乙醯基轉移??的特性以及其對於魚類多胺類與能量代謝的影響
Using Zebra Fish as a Model to Study the Properties of Spermidine/Spermine Acetyltransferase I (Ssat1) and Evaluate Its Influences on Polyamine as Well as Energy Metabolism in Fish
Authors: 林翰佳;胡清華
Contributors: NTOU:Institute of Bioscience and Biotechnology
Date: 2011-08
Issue Date: 2012-04-13T01:06:33Z
Publisher: 行政院國家科學委員會
Abstract: 摘要:多胺 (polyamine, PA) 泛指腐胺 (Put)、亞精胺 (Spd) 以及精胺 (Spm), 廣泛地存在於各物種中, 是細胞生存的必需因子。而 SSAT1 (Spermidine/Spermine AcetylTransferase 1) 是高等動物所獨有的PA 代謝酵 素,可將乙醯基從 acetyl-CoA 上轉移到 Spd 或 Spm 上,因而促進 PA 的 降解或向胞外運送。由於 SSAT1 是代謝 PA 的關鍵的酵素,因此基因表 現受到嚴格調控。其中最特殊的現象是該基因 ORF 區域會被一種 RNA 結合蛋白控制,因此只有在高濃度 PA 之下才能轉譯 SSAT1 蛋白質。由 最近的研究顯示SSAT1 除了參與 PA 代謝之外,也可以直接結合其他蛋 白質而參與相關訊息傳導路徑,因而被認為是許多生理調控途徑的關鍵因 子。 本團隊發現在斑馬魚中有 3 個與 SSAT 相似的基因 (zSSAT1, zSSAT2 與 zDAT),並已初步鑑定部分的酵素活性與轉譯調控特性。雖然 其中的 zSSAT1 與 zDAT 兩者的胺基酸有 80% 相同,但在蛋白質轉譯特 性上,zDAT 與人類 SSAT1 同樣受 PA 調控,而zSSAT1 則沒有。從酵 素基質選擇性來看 zSSAT1 也與人類酵素有所差異。由我們的發現顯示, 這些特性各異的基因可以成為研究 SSAT1 功能的最佳範本。在本計劃 中,我們將先針對斑馬魚 SSAT 相關基因的酵素特性、調控機制以及與其 他蛋白相互作用等特性進行全面的比較。再以突變的方式探討相關基因序 列的差異對於酵素活性、調控機制以及與其他蛋白相互作用等方面造成的 影響。透過酵素特性的掌握,我們將可以建立斑馬魚細胞內 SSAT1 活性 過量或抑制的基因轉殖模式,並利用這些模式研究 SSAT 對斑馬魚能量代 謝所造成的影響。
Abstract:Polyamines, including putrescine, spermidine and spermine, are vital factors for cells and widely distributed in nearly all kinds of species. Spermidine/Spermine Acetyl Transferase 1 (SSAT1), a polyamine catabolic enzyme exclusively expressed in higher animals, transferring an acetyl group from acetyl-CoA to polyamine, facilitates polyamines’ degradation or exportation. Being the key enzyme in the polyamine catabolism pathway, the expression of SSAT1 is strictly regulated, especially at translation level. The ORF region of SSAT1 mRNA is restricted by an unknown protein, which will release its inhibition of ribosome when cellular polyamine increased. In addition to its role on polyamine catabolism, recent studies indicated that SSAT1 may directly interact with other proteins and involving with other physiological events. Therefore, SSAT1 is considered as a key regulator in many aspects. Previously, we identified 3 SSAT-like genes (denoted as zSSAT1, zSSAT2 and zDAT) from zebrafish. From our preliminary studies, we found that only zDAT but not zSSAT1 displays similar translational regulatory profile as human gene. Furthermore, the catalysis property of zSSAT1 is also different from human SSAT1. Despite of their overall sequence identity is higher than 75%, these genes display different characteristics. Thus they may be good templates to study the structure-function relationship of SSAT1. In this project, we would like to characterize the enzyme activity, mechanism of gene regulation and protein-protein interaction of all three genes. Further, mutant enzymes will be prepared to elucidate the important domains for enzyme activity and protein interaction. In addition to the in vitro studies, functions of these genes are also examined on cell or animal models. Gain-of-function or loss-of-function transgenic zebrafish models will be used to evaluate the influence of SSAT1 activity on metabolism.
Relation: NSC99-2313-B019-010-MY3
Appears in Collections:[Department of Bioscience and Biotechnology ] Research Reports

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