The 3a accessory protein of SARS coronavirus specifically interacts with the 5'UTR of its genomic RNA, Using a unique 75 amino acid interaction domain.
The 3a accessory protein of SARS coronavirus specifically interacts with the 5'UTR of its genomic RNA, Using a unique 75 amino acid interaction domain.
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DOI：
10.1021/bi062057p
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发表时间：
2007-06
期刊：
影响因子：
2.9
通讯作者：
K. Sharma;Milan Surjit;N. Satija;Bo-ping Liu;V. Chow;S. Lal
中科院分区：
文献类型：
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作者：
K. Sharma;Milan Surjit;N. Satija;Bo-ping Liu;V. Chow;S. Lal
严重急性呼吸道综合征（SARS）流行病爆发以来已经过去了四年多，但人们对这种致命病毒的分子生物学和发病机制仍然知之甚少。在SARS冠状病毒（SARS-CoV）的辅助蛋白中，3a蛋白已被证明与刺突、包膜和膜糖蛋白相互作用，并且最近被确定为衣壳的结构组分。最近的研究表明，3a蛋白可能作为一个离子通道，并可能促进病毒的释放。为了进一步表征该蛋白的功能特性，我们开始研究以检查其RNA结合活性。利用酵母三杂交系统、电泳迁移率变动分析（EMSA）和紫外交联技术，我们证明了3a蛋白能够与SARS病毒基因组RNA的5'非翻译区（5' UTR）特异性结合。此外，我们已经映射的相互作用结构域的3a蛋白负责这种RNA-蛋白质的相互作用，使用一系列的缺失突变体，并将其定义为中央75个氨基酸的区域。3a的这种RNA结合基序与任何其他已知的RNA结合蛋白不具有同源性，并且可能在病毒衣壳组装和发病机制中具有重要作用。
More than four years have passed since the outbreak of the severe acute respiratory syndrome (SARS) epidemic, and still very little is known about the molecular biology and pathogenesis of this deadly virus. Among the accessory proteins of the SARS coronavirus (SARS-CoV), the 3a protein has been shown to interact with the spike, envelope, and membrane glycoprotein and has recently been established to be a structural component of capsid. Recent studies suggest that the 3a protein may function as an ion channel and may promote virus release. In order to further characterize the functional properties of this protein, we initiated studies to check its RNA binding activity. Using the yeast three-hybrid system, electrophoretic mobility shift assay (EMSA), and ultraviolet (UV) cross-linking techniques, we have shown that the 3a protein is capable of binding specifically to the 5' untranslated region (5'UTR) of the SARS virus genomic RNA. Further, we have mapped the interaction domain of the 3a protein responsible for this RNA-protein interaction using a series of deletion mutants and defined it to the central 75 amino acid region. This RNA binding motif of 3a does not share homology with any other known RNA binding protein and may have an important role in viral capsid assembly and pathogenesis.