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Revealing the HIV-1 Interactome

揭示 HIV-1 相互作用组

基本信息

批准号：
8415922
负责人：
MARK AYER MUESING
金额：
$ 42.03万
依托单位：
AARON DIAMOND AIDS RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-15 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415922
关键词：
Affinity Antigens Antiviral Agents Binding Biochemical Biological Biological Preservation Biological Process Cell physiology Cells Code Complex DNA DNA Viruses Engineering Environment Enzymes Epitopes Funding Gene Targeting Genetic Genetic Code Genome Growth HIV HIV Infections HIV-1 Health Human Human Activities Infection Integrase Integration Host Factors Intervention Investigation Life Cycle Stages Maps Mass Spectrum Analysis Mediating Methodology Methods Mutagenesis Nuclear Import Procedures Process Proteins Proteome Proteomics Protocols documentation Reverse Transcription Site Site-Directed Mutagenesis Source Structure-Activity Relationship Surface Surveys System T-Lymphocyte Techniques Ubiquitination Vaccines Viral Viral Envelope Proteins Viral Genome Viral Proteins Virion Virus base cryogenics env Gene Products immunological intervention insight novel protein complex protein protein interaction recombinant virus small molecule sound tool vif Gene Products viral DNA

项目摘要

DESCRIPTION (provided by applicant): Given the limited genetic coding capacity of HIV-1, it is reasonable to expect that the virus must interact with an extensive battery of cellular factors to complete its passage through the cell. Indeed, it is remarkable that the viral genome, comprising only about 0.0003% of the entire genetic capacity of the cell, commandeers the cellular environment to its own advantage. However, to date and despite being easily the most studied virus, only a relatively small group of cellular proteins have been shown essential for HIV-1 propagation. A genetic/proteomic approach has been developed that utilizes a random mutagenic protocol followed by a stringent step of biological selection in efforts to pinpoint those rare sites within the HIV-1 proteome that can accommodate the incorporation of a potent affinity tag without significant loss of those structure/function relationships that are required for viral replication-competency. In conjunction with a novel cryogenic methodology to capture and preserve transient viral-host interactions during immunoisolation, we have recovered several host proteins in association with the replicating virus but previously obscured from conventional scientific investigation. We are reassured that our method is sound since among the cellular proteins recovered and identified using our methodologies are those previously determined by other groups to interact with HIV-1. Furthermore, a straightforward mass spectrometric technique is presented that discriminates with high accuracy between specific and nonspecific proteins in immunoisolated protein complexes. The combined genetic and proteomic method described is a powerful tool set that can be utilized to identify cellular proteins in transient association with a given viral protein at points during the viral life cycle, a subset of which may be absolutely required by the virus for its livelihood but in part, dispensable by the host cell.

描述（由申请人提供）：鉴于HIV-1的遗传编码能力有限，可以合理预期病毒必须与大量细胞因子相互作用才能完成其通过细胞的过程。事实上，值得注意的是，病毒基因组只占细胞全部遗传能力的0.0003%，却能利用细胞环境为自己谋利。然而，迄今为止，尽管是最容易被研究的病毒，但只有相对较少的一组细胞蛋白质被证明是HIV-1繁殖所必需的。已经开发了一种遗传/蛋白质组学方法，该方法利用随机诱变方案，然后进行严格的生物选择步骤，以努力查明HIV-1蛋白质组内的那些罕见位点，这些位点可以容纳有效的亲和标签的掺入，而不会显著损失病毒复制能力所需的那些结构/功能关系。结合一种新的低温方法来捕获和保存免疫隔离过程中短暂的病毒-宿主相互作用，我们已经恢复了几个与复制病毒相关的宿主蛋白，但以前从传统的科学调查中被掩盖。我们确信，我们的方法是合理的，因为在使用我们的方法回收和鉴定的细胞蛋白质中，有那些先前由其他小组确定与HIV-1相互作用的蛋白质。此外，一个简单的质谱技术，提出了高精度的特异性和非特异性蛋白质之间的免疫分离的蛋白质复合物的歧视。所描述的组合的遗传和蛋白质组学方法是一个强大的工具集，其可用于鉴定在病毒生命周期期间的点处与给定病毒蛋白质瞬时相关的细胞蛋白质，其中的一个子集可能是病毒对其生计绝对需要的，但部分是由宿主细胞替代的。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Suppression of Foxo1 activity and down-modulation of CD62L (L-selectin) in HIV-1 infected resting CD4 T cells.

DOI：
10.1371/journal.pone.0110719
发表时间：
2014
期刊：
PloS one
影响因子：
3.7
作者：
Trinité B;Chan CN;Lee CS;Mahajan S;Luo Y;Muesing MA;Folkvord JM;Pham M;Connick E;Levy DN
通讯作者：
Levy DN

Mass spectrometry-based proteomic approaches for discovery of HIV-host interactions.

基于质谱的蛋白质组学方法用于发现 HIV-宿主相互作用。