Crystallographic studies of retroviral intasome complexes

逆转录病毒嵌体复合物的晶体学研究

基本信息

批准号：
8919420
负责人：
Hideki Aihara
金额：
$ 41.82万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8919420
关键词：
Acquired Immunodeficiency Syndrome Address Anti-HIV Agents Antiviral Agents Architecture Binding Biological Models Cells Chromatin Chromosomes Complex Cryoelectron Microscopy Crystallization DNA DNA Binding Domain Detergents Drug resistance Enzymes Genome Genomic DNA Goals HIV-1 Health Integrase Integrase Inhibitors Label Left Length Light Mercury Methods Molecular Chaperones Mutagenesis Mutation Nucleoproteins Phase Protein Engineering Proteins Reaction Research Resolution Retroviridae Roentgen Rays Rous sarcoma virus Solubility Spumavirus Structure Subfamily lentivirinae System Techniques Testing Vertebral column Viral Visna-maedi virus X-Ray Crystallography base cellular targeting clinically relevant design genetic information improved inhibitor/antagonist insight lens epithelium-derived growth factor next generation novel strategies particle prototype resistance mechanism success transcriptional coactivator p75 viral DNA

Acquired Immunodeficiency Syndrome Address Anti-HIV Agents Antiviral Agents Architecture Binding Biological Models Cells Chromatin Chromosomes Complex Cryoelectron Microscopy Crystallization DNA DNA Binding Domain Detergents Drug resistance Enzymes Genome Genomic DNA Goals HIV-1 Health Integrase Integrase Inhibitors Label Left Length Light Mercury Methods Molecular Chaperones Mutagenesis Mutation Nucleoproteins Phase Protein Engineering Proteins Reaction Research Resolution Retroviridae Roentgen Rays Rous sarcoma virus Solubility Spumavirus Structure Subfamily lentivirinae System Techniques Testing Vertebral column Viral Visna-maedi virus X-Ray Crystallography base cellular targeting clinically relevant design genetic information improved inhibitor/antagonist insight lens epithelium-derived growth factor next generation novel strategies particle prototype resistance mechanism success transcriptional coactivator p75 viral DNA

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项目摘要

DESCRIPTION (provided by applicant): Retrovirus integration is catalyzed by the viral integrase (IN), which forms a tetrameric complex with both ends of the linear viral DNA genome and captures a cellular target DNA for concerted insertions of the viral DNA termini. The IN-DNA complex formed by HIV-1 IN is the target of a class of antiviral drugs called IN strand- transfer inhibitors. A better understanding of the IN-DNA interactions is therefore important for improving the clinically relevant anti-HIV drugs as well as designing next-generation IN inhibitors. The recent success in the structural studies of prototype foamy virus (PFV) IN-DNA complexes led to tremendous amount of insights into the retrovirus integration reaction. However, structural characterization of IN-DNA complexes from any other retrovirus systems by X-ray crystallography has remained elusive, leaving open the question as to how well the architecture of the IN-DNA complexes is conserved between distantly related retrovirus systems. We hypothesize that domain arrangement and IN-DNA interactions within the intasome, a nucleoprotein complex containing the tetramer of IN with the viral and target DNA molecules, are different between the canonical 3- domain IN including HIV-1 IN and the larger 4-domain PFV IN featuring an additional DNA-binding domain and longer inter-domain linkers. To address this hypothesis, we will determine crystal structures of the intasome complexes formed by the canonical 3-domain retroviral INs. Using a novel approach we have developed, crystals of the Rous sarcoma virus intasome suitable for structure determination have been obtained. We will further use our technique to pursue the crystal structure of a lentiviral intasome bound to the host co-factor LEDGF/p75. Our X-ray crystallographic analyses will provide the critically needed structural information to better understand the integration of HIV-1 and closely related retroviruses.

描述（由申请人提供）：逆转录病毒的整合被病毒整合酶（IN）催化，该酶形成了具有线性病毒DNA基因组的两端的四聚体络合物，并捕获了一个细胞靶DNA，以用于病毒DNA末端的协同插入。 HIV-1在IN中形成的DNA复合物是在链转移抑制剂中称为一类抗病毒药的靶标。因此，更好地了解DNA相互作用对于改善临床相关的抗HIV药物以及在抑制剂中设计下一代很重要。在原型泡沫病毒（PFV）内DNA复合物的结构研究中，最近的成功导致了对逆转录病毒整合反应的大量见解。然而，通过X射线晶体学对任何其他逆转录病毒系统中DNA复合物的结构表征仍然难以捉摸，这留下了一个问题，即DNA复合物的体系结构在远距离相关的逆转录病毒系统之间的保守程度。我们假设在该内体内的结构域排列和DNA相互作用，即含有病毒和靶DNA分子IN的核蛋白蛋白复合物，在包括HIV-1 IN（包括HIV-1）和较大的4-DOMAIN PFV中，在较大的4-DOMAIN PFV中具有附加的DNA结合结构域和更长的较长的较长层层次链接。为了解决这一假设，我们将确定由规范3域逆转录病毒INS形成的内塔体复合物的晶体结构。使用一种新的方法，我们已经获得了适合结构测定的Rous Sarma病毒Intasome的晶体。我们将进一步使用我们的技术来追求与宿主副因子LEDGF/p75结合的慢病毒Intasome的晶体结构。我们的X射线晶体学分析将提供迫切需要的结构信息，以更好地了解HIV-1和密切相关的逆转录病毒的整合。