CONTROL OF HIV REPLICATION BY CYCLIN L2

细胞周期蛋白 L2 对 HIV 复制的控制

基本信息

批准号：
9089866
负责人：
George Kyei
金额：
$ 16.45万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-15 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9089866
关键词：
Acquired Immunodeficiency Syndrome Adverse drug effect Affect Alternative Splicing Amino Acids Anti-Retroviral Agents Arginine Attention Biochemical Boxing C-terminal Cell Cycle Cell Cycle Regulation Cells Cellular biology Complex Cyclins Data Dendritic Cells Fatigue Genes Genome Genomics Goals HIV HIV-1 HIV-2 Hybrids Immune Immunoprecipitation Infection Interphase Cell Interruption K-Series Research Career Programs Knowledge Lead Life Light Literature Maps Mass Spectrum Analysis Mentors Mentorship Messenger RNA Methods Molecular Molecular Biology Molecular Genetics Molecular Target N-terminal Patients Phosphorylation Phosphotransferases Physicians Polymerase Chain Reaction Positioning Attribute Process Proteins Proteomics Quality of life Quantitative Reverse Transcriptase PCR RNA Splicing Resources Rest Role SIV Scientist Serine Skills Development Source T-Lymphocyte Testing Training Transcript United States Universities Viral Viral Load result Viral reservoir Virus Virus Replication Washington Work Yeasts antiretroviral therapy attenuation base cell type cost experience improved innovation kinase inhibitor knock-down mRNA Precursor macrophage molecular domain multicatalytic endopeptidase complex new therapeutic target novel novel therapeutics pandemic disease public health relevance research study skill acquisition therapeutic target transcriptome sequencing ubiquitin ligase whole genome

项目摘要

DESCRIPTION (provided by applicant): The goal of this Mentored Career Development Award is to facilitate Dr. George Kyei's transition to independence as a physician-scientist studying how the human immunodeficiency virus (HIV) is able to survive in latently infected cell like resting T cells and macrophages. The proposed experiments will be conducted under the mentorship of Dr. Lee Ratner, and they will examine the role of the protein cyclin L2 and how it controls HIV replication in the aforementioned cell types. HIV continues to be a global pandemic with over 35 million people living with the virus. In the United States, over 50,000 new infections happen every year. Although current anti-retroviral therapy (ART) can suppress the virus and improve the quality of life for patients, these treatments do not lead to cure. Within weeks of patients stopping therapy, the viral load begins to rise again. Lifelong treatment has the attendant problems of patient fatigue, drug side effects and unsustainable costs. Hence viral elimination or cure is a vital goal. The main reason for viral resurgence upon treatment interruption is the presence of viral reservoirs like resting T cell and macrophages, which have the virus integrated in their genomes in a resting state. As soon as treatment is interrupted, these cells serve as sources of virus. Despite their crucial role in sustaining HIV persistence, current knowledge of viral replication requirements in macrophages and resting T cells is very limited. Recently, we identified cyclin L2 as critical for HIV replication in macrophages. The goal of this project is to test the hypothesis that cyclin L2 controls HIV replication in macrophages and resting T cells by modulating viral alternative splicing in cooperation with specific cellular kinases. In order to test this hypothesis, we will use biochemical, molecular genetics, genomics and proteomics approaches to test the following specific aims: 1. to characterize the mechanisms by which cyclin L2 controls HIV pre-mRNA alternative splicing. We will use gene knockdown, polymerase chain reaction, whole genome RNA sequencing and biochemical methods to define how cyclin L2 controls the splicing of HIV. 2. To determine the effect of phosphorylation on cyclin L2 function. Cyclin L2 is known to be heavily phosphorylated. We will use proteomics, gene knockdown approaches and specific kinase inhibitors to investigate the importance of Dyrk1A and CDK11B as potential kinases for cyclin L2. 3. To identify and characterize the molecular domains of cyclin L2 required supporting HIV replication in non- cycling cells. The rationale for this aim is to identify the exact motifs of cyclin L2 required forHIV replication, in order to map out a potential therapeutic target. . The proposed experiments will shed new light in our understanding of HIV replication requirements in these critical cell types and potentially reveal novel molecular targets for HIV elimination.

描述：指导发展奖的目的是促进CE博士作为医生 - 科学家研究人类免疫缺陷病毒（HIV）如何能够在静止的T细胞（例如静止的T细胞和巨噬细胞）中生存。在美国有超过3500万人的病毒中，进行了指导Lee Ratner Clin L2及其如何控制艾滋病毒。每年都会在几周内停止治疗对于治疗中断时的病毒率是静止的T细胞和巨噬细胞的保留，这些病毒在其基因组中的静止状态很快持续性HIV持续性，最近对巨噬细胞和静息T细胞中病毒复制要求的知识受到限制，我们确定Cyclin L2对于巨噬细胞中的HIV复制至关重要。该项目的假设是，Cyclin L2通过与特定的细胞激酶合作，控制巨噬细胞和静止的T细胞中的HIV复制。使用基因敲低，聚合酶链反应，整个基因组RNA测序和生化方法来定义Cyclin L2如何控制HIV的剪接。 dyrk1a和cdk11b作为细胞周期蛋白L2的潜在激酶。在我们对HIV D的新发现中，有可能复兴的新型分子靶标，以消除HIB。