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標題: 從分子層次探討人類冠狀病毒(229E與OC43)和A型流感病毒(H1N1)核殼蛋白與核酸之結合機制
Mechanistic Study of Nucleocapsid Proteins of Human Coronavirus (229E and OC43) and Influenza Avirus (H1N1) Bound to Nucleic Acids at Molecular Level
作者: 侯明宏
關鍵字: 生物科學類;基礎研究
人類冠狀病毒229E、OC43 與A 型流感病毒H1N1 在寒冷的季節當中,主要會引起人類普通感冒與流行性感冒的發生。由於普通感冒與流行性感冒所造成的症狀相當類似,症狀亦難以區分。一般而言,普通感冒所引起的症狀會比流感還要溫和。然而,由於病毒多突變特性,讓冠狀病毒229E、OC43 和A 型流感病毒H1N1 可能對於台灣人民的健康會造成很大的影響,因此,致力於此兩類病毒的研究可以說是刻不容緩。在病毒當中,內部的核殼蛋白(nucleocapsid protein) 主要功能是與RNA 核酸單鏈結合,並組裝形成螺旋狀的核醣蛋白質複合體(RNP),核醣蛋白質複合體形成的目的主要為了能夠維持病毒RNA 在恆定的構形下複製與轉錄,對於病毒的繁殖可以說是相當重要。而到目前為止,這兩類病毒利用核殼蛋白組裝螺旋的機制尚不是十分清楚。在先前的計畫當中,我們已經利用若干生化和生物物理的方法來研究人類冠狀病毒229E 與OC43 核殼蛋白的生化特性和功能區域(domain)。延續先前的計畫,未來三年的計劃當中主要是針對人類冠狀病毒229E、OC43和A 型流感病毒H1N1 與核酸結合的分子機制所進行之重要研究。以下五點為本計劃的重要方向及執行的時程:1. 分析冠狀病毒229E、OC43 與A 型流感病毒H1N1 核殼蛋白所結合之核醣核酸序列專一性與其之間的動力學行為。2. 確認冠狀病毒229E 和OC43 核殼蛋白與核醣核酸結合之重要胺基酸。3. 分析A 型流感病毒H1N1 核殼蛋白之摺疊和穩定性。4. 確認A 型流感病毒H1N1 核殼蛋白與核醣核酸結合區域和重要胺基酸5. 分析冠狀病毒229E 及OC43 和A 型流感病毒H1N1 核殼蛋白與核醣核酸結合之晶體結構。本實驗室主要是針對病毒之核殼蛋白與核酸結合之分子機制來進行一些重要的研究。到目前為止,我們也已經獲得一些成果。因此計劃的結果也將會對於發展台灣研發抗病毒藥物、疫苗與其診斷方法平台也會有相當大的助益,未來我們亦會慢慢朝向這些方向來進行。

Common colds and influenza caused by human coronaviruses (HCoV) and influenza A virusrespectively, are both highly contagious diseases in the colder months. Ii is quit difficult to distinguishthese two diseases due to their similar symptoms, however. In general, common colds have a propensityto be mild, compared with flu. Nucleocapsid (N) protein, the major virion structural protein, has beenidentified to bind with the viral genomic RNA and form ribonucleocapsid (RNP) in the viruses. Theformation of the RNP may also be important in terms of maintaining the RNA in an ordered conformationsuitable for replication and transcription of the viral genome. Nevertheless, in HCoV and influenzavirus, the molecular mechanism of helical packing of RNP remains unclear. Our previous study usingbiochemical and biophysical methods revealed the biochemical properties and domain organization ofHCoV-OC43 and 229E N protein. By following with our previous studies, we are interested inunderstanding the molecular mechanism of RNA packaging in nucleocapsid proteins of HCoV (229E andOC43) and influenza virus A. Here, we have designed a three-years project that is mainly aiming at themechanistic study of nucleocapsid protein of HCoV-OC43 and 229E and influenza A virus (H1N1) boundto nucleic acids at molecular level. There are five goals with plan implementation in this proposal listedas follows.1. Characterization of sequence-specific binding and kinetic behavior of HCoV-229E,HCoV-OC43, and influenza A virus (H1N1) nucleocapsid proteins to single-stranded RNA (orDNA).2. Identification of crucial amino acids of HCoV-229E and HCoV-OC43 nucleocapsid proteinsthat function as RNA-binding elements.3. Study of folding and stability of influenza A virus (H1N1) nucleocapsid protein.4. Identification of RNA-binding domain and crucial amino acids of influenza A virus (H1N1)nucleocapsid protein.5. Structural analyses of HCoV-229E, HCoV-OC43, and influenza A virus (H1N1) nucleocapsidproteins bound to RNA.Until now, several preliminary data have been finished and presented in this proposal. This studyis expected to benefit the development of drugs capable of disrupting the binding of viral N protein withRNA and in turn viral assembly in the future.
其他識別: NSC97-2311-B005-003-MY3
Appears in Collections:生物科技發展中心

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