HIV-1 Preintegration Trafficking and Nuclear Localization

HIV-1 融入社会前贩运和核定位

• 批准号: 7767733
• 负责人: Alan N. Engelman
• 金额: $ 41.52万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-15 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7767733
• 关键词: AIDS/HIV problem; Acute; Address; Affect; Behavior; Biochemical; Biochemistry; C-terminal; Capsid; Cell Nucleus; Cells; Cellular biology; Chimeric Proteins; Chromatin; Chromosomes; Collaborations; Complementary DNA; Complex; Confocal Microscopy; Cytoplasm; DNA; DNA biosynthesis; Defect; Doctor of Philosophy; Drug Delivery Systems; Dynein ATPase; Fluorescent in Situ Hybridization; Fractionation; Funding; Fv-1 protein; Gagging; Goals; Grant; HIV-1; Hela Cells; Human; Importins; In Vitro; Infection; Integrase; Integration Host Factors; Interphase Cell; Knock-out; Knowledge; MG132; Macaca mulatta; Mediating; Methods; Microtubules; Minor; Modeling; Molecular; Murine leukemia virus; Mutation; Nuclear; Nuclear Import; Nuclear Protein; Nuclear Proteins; Nucleic Acids; Pharmaceutical Preparations; Phenotype; Process; Proteasome Inhibitor; Proteins; Proteomics; RNA Interference; Reporting; Research Personnel; Reverse Transcription; Role; Running; Science; Techniques; Therapeutic Intervention; Update; Viral; Virus; Virus Diseases; base; emerin; fight against; macrophage; monocyte; mutant; new therapeutic target; novel; research study; theories; trafficking; transcriptional coactivator p75; vector; virus host interaction

DESCRIPTION (provided by applicant): The most effective approach for managing HIV/AIDS is through therapeutic intervention. Current anti-viral drugs target reverse transcription, virus maturation, and entry. Novel drugs against as of yet unexploited targets are essential and any replication step that the virus must accomplish to grow in human cells defines a viable target. After entering a susceptible target cell, the virus must move from the cell periphery, through the cytoplasm, and into the nucleus where it will recombine its neo- synthesized DNA with a cell chromosome. Mutations that impede preintegration trafficking are lethal to the virus, yet the mechanistic details of how the virus moves through the cytoplasm and enters the nucleus are poorly understood. This proposal will uncover essential mechanistic details of HIV-1 preintegration trafficking and nuclear import using a tour de force of molecular techniques. Proteomic approaches will be utilized to uncover essential virus-host interactions. Biochemical fractionation will be utilized to compare the composition of wild-type and mutant replication complexes, and fluorescent confocal microscopy will be used to visualize the complexes as they move through the cell. Fluorescent in situ hybridization will be used to visualize viral nucleic acids once they are inside the nucleus. The results of these experiments will reveal salient features of how the virus moves through the cell to accomplish the essential preintegration tasks in its lifecycle. This knowledge in the long run will define novel targets for therapeutic interaction to develop new anti-viral drugs in the fight against HIV/AIDS.

描述（由申请人提供）： 管理艾滋病毒/艾滋病的最有效方法是通过治疗干预。目前的抗病毒药物以逆转录、病毒成熟和进入为目标。针对尚未开发的靶标的新药至关重要，病毒在人体细胞中生长必须完成的任何复制步骤都定义了可行的靶标。进入易感靶细胞后，病毒必须从细胞外周移动，穿过细胞质，进入细胞核，在那里它将新合成的 DNA 与细胞染色体重新组合。阻碍整合前运输的突变对病毒来说是致命的，但人们对病毒如何穿过细胞质并进入细胞核的机制细节知之甚少。该提案将利用分子技术的杰作，揭示 HIV-1 整合前贩运和核进口的基本机制细节。将利用蛋白质组学方法来揭示重要的病毒与宿主的相互作用。将利用生化分级分离来比较野生型和突变型复制复合物的组成，并使用荧光共聚焦显微镜观察复合物在细胞中移动时的情况。一旦病毒核酸进入细胞核，荧光原位杂交将用于可视化病毒核酸。这些实验的结果将揭示病毒如何穿过细胞以完成其生命周期中基本的预整合任务的显着特征。从长远来看，这些知识将为治疗相互作用确定新的目标，以开发新的抗病毒药物来对抗艾滋病毒/艾滋病。