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Dynamic Interplay of RNA and Protein in the Human Immunodeficiency Virus-1 Reverse Transcription Initiation Complex

机译:RNA和蛋白质在人免疫缺陷病毒-1逆转录络合物中的动态相互作用

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The initiation of reverse transcription in human immunodeficiency virus-1 is a key early step in the virus replication cycle. During this process, the viral enzyme reverse transcriptase (RT) copies the single-stranded viral RNA (vRNA) genome into double-stranded DNA using human tRNALYs3 as a primer for initiation. The tRNA primer and vRNA genome contain several complementary sequences that are important for regulating reverse transcription initiation kinetics. Using single-molecule Forster resonance energy transfer spectros-copy, we demonstrate that the vRNA tRNA initiation complex is conformationally heterogeneous and dynamic in the absence of RT. As shown previously, nucleic acid RT interaction is characterized by rapid dissociation constants. We show that extension of the vRNA tRNA primer binding site helix from 18 base pairs to 22 base pairs stabilizes RT binding to the complex and that the tRNA 5' end has a role in modulating RT binding. RT occupancy on the complex stabilizes helix 1 formation and reduces global structural heterogeneity. The stabilization of helix 1 upon RT binding may serve to destabilize helix 2, the first pause site for RT during initiation, during later steps of reverse transcription initiation. (C) 2018 Elsevier Ltd. All rights reserved.
机译:反转录在人免疫缺陷病毒1的启动是在病毒复制周期的关键的早期步骤。在此过程中,病毒逆转录酶(RT)复制单链病毒RNA(vRNA的)基因组中双链DNA使用人tRNALYs3作为起始引物。 tRNA引物和基因组vRNA的包含可用于调节反向转录起始动力学重要几个互补的序列。使用单分子福斯特共振能量转移spectros拷贝,我们证明了的vRNA tRNA的起始复合物是在没有RT的构象异构和动态的。如前所示,核酸RT相互作用的特征在于快速解离常数。我们表明,延长的vRNA的tRNA引物结合位点的螺旋从18个碱基对至22个碱基对稳定RT结合于复杂和tRNA的5' 端具有在调节RT结合的作用。在复企稳RT占用螺旋1层的形成和减少全球结构异质性。螺旋1的在RT稳定结合可以用于动摇螺旋2,开始在所述第一暂停网站RT,在反转录起始的后面的步骤。 (c)2018年elestvier有限公司保留所有权利。

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