权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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

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（由申请方提供）：逆转录病毒整合由病毒整合酶（IN）催化，其与线性病毒DNA基因组的两端形成四聚体复合物，并捕获细胞靶DNA以协同插入病毒DNA末端。由HIV-1 IN形成的IN-DNA复合物是一类称为IN链转移抑制剂的抗病毒药物的靶标。因此，更好地理解IN-DNA相互作用对于改进临床相关的抗HIV药物以及设计下一代IN抑制剂是重要的。最近在原型泡沫病毒（PFV）IN-DNA复合物的结构研究中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射线晶体学对来自任何其他逆转录病毒系统的IN-DNA复合物的结构表征仍然是难以捉摸的，留下了关于IN-DNA复合物的结构在远亲逆转录病毒系统之间如何保守的问题。我们假设，结构域的安排和IN-DNA相互作用内的整合体，一个核蛋白复合物含有四聚体的IN与病毒和靶DNA分子，是不同的典型的3-域IN，包括HIV-1 IN和更大的4-域PFV IN具有额外的DNA结合域和更长的域间连接。为了解决这一假设，我们将确定晶体结构的整合体复合物形成的典型的3-域逆转录病毒INs. Using一种新的方法，我们已经开发出，晶体的劳斯肉瘤病毒整合体适合于结构测定。我们将进一步使用我们的技术来研究与宿主辅因子LEDGF/p75结合的慢病毒整合体的晶体结构。我们的X射线晶体学分析将提供急需的结构信息，以更好地了解HIV-1和密切相关的逆转录病毒的整合。