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    题名: 利用人類蛋白質體微陣列晶片探究病毒核醣核酸與宿主之交互作用;Deciphering the viral RNA-host interactions using human proteome microarrays
    作者: 陳健生;吳淩嫣
    贡献者: 國立中央大學系統生物與生物資訊研究所
    关键词: 生物科學;Proteme chip;RNA virus;Virus-host interaction
    日期: 2015-01-08
    上传时间: 2015-01-13 12:10:53 (UTC+8)
    出版者: 財團法人國家衛生研究院
    摘要: 研究期間:10301~10312;Over the past few decades, the incidence and prevalence of diseases caused from RNA viruses is becoming more frequent and serious, bringing a critical health concern all over the world. Basically, a successful infection by the virus requires proper interactions between viral RNA molecules and cellular proteins in the host. Among the interactions, the untranslated regions (UTRs) of virus RNA genome play a pivotal role in genome accumulation and virion production. Thus, deciphering of the protein-viral UTR interactions allows a better understanding of those pathogenic mechanisms and provides potential therapeutic targets for developing antivirals. However, the most considerable challenge in the study of protein-RNA interactions is that testing a singular interaction at a time is cumbersome and not practical. To study the interactions in a high-throughput manner, an array of ~17,000 non-redundant human proteins has been recently established in our collaborative team. Herein, we intend to implement the comprehensive proteome chip analysis to identify proteins that recognize the conserved UTRs of hepatitis C virus (HCV). In addition, it has been implicated that microRNA-122 (miR-122) participates in virion production and stimulation of virus ranslation through binding to its target sequence on HCV 5’ UTR. To globally identify the functional roles of miR-122 in the context of HCV infection, profiling of the miR-122 interacting proteins will be carried out using the human proteome chip. Proteins with binding capacities to both SL1 and miR-122 might be directly involved in HCV biology. After identifying HCV UTR-binding proteins, those proteins will be high-throughput purified and spotted on aldehyde slides to construct the “boutique” chips. Compared to the human proteome chip, this boutique chip will be easier to fabricate at lower cost, and will allow us to efficiently identify the interaction inhibitors in the future. To demonstrate the utility of the novel screening platform, a cohort of FDA approved NA-binding antibiotics will be used to probe the boutique chips. The antibiotic that sequesters the protein-viral UTR interactions could be a potential antiviral drug against HCV infection. All the biological significance of identified interactions and the efficacy of inhibitors will be further studied to complete our aims. Remarkably, our team has recently identified a protein called hnRNP K which specifically binds to the tetraloop and the miR-122 seed sequence in the stem loop 1 (SL1) of HCV 5’ UTR. We showed that hnRNP K knockdown contributes to HCV RNA decreased by approximately 4-fold. These current evidences indicate that hnRNP K served as a chaperon that directs miR-122 to bind HCV SL1 and enhance HCV genome replication. Obviously, our preliminary data strongly supports the feasibility and high impact of the proposed project.
    關聯: 財團法人國家實驗研究院科技政策研究與資訊中心
    显示于类别:[系統生物與生物資訊研究所] 研究計畫

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