Cellular Determinants and Function Consequences of PP2A-B56 Degradation by HIV-1 Vif

HIV-1 Vif 降解 PP2A-B56 的细胞决定因素和功能后果

基本信息

批准号：
10619722
负责人：
Jeffrey R Johnson
金额：
$ 55.14万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-05 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10619722
关键词：
APOCEC3G gene Acquired Immunodeficiency Syndrome Address Affinity Chromatography Binding Biological Assay CD4 Positive T Lymphocytes CRISPR screen CRISPR/Cas technology Cell Line Cells Chronic Data Development Drug Targeting Environment Event Family Flow Cytometry Fluorescence Gene Silencing Genes Genetic Screening HIV-1 Holoenzymes Human Individual Infection Inflammation Inflammatory Label Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Metabolic Diseases Molecular Molecular Virology Pathway interactions Persons Phosphorylation Phosphorylation Site Play Process Protein Family Protein Subunits Protein phosphatase Proteins Proteomics Resources Retroviridae Role Signal Transduction Site Technology Testing Translations Ubiquitination Viral Viral Physiology Virion Work antiretroviral therapy base editing cDNA Expression candidate identification cardiovascular disorder risk community transmission drug development engineered T cells fitness genome-wide multidisciplinary novel phosphoproteomics prevent protein complex protein degradation reactivation from latency targeted treatment therapy development vif Gene Products virology virus host interaction

项目摘要

PROJECT SUMMARY The HIV-1 Vif protein is expressed late during infection and has a well-described function to ubiquitinate and degrade proteins in the APOBEC3 family, thus neutralizing their antiviral activities. More recently, an additional function of HIV-1 Vif was described by us and others to ubiquitinate and degrade protein phosphatase 2A holoenzymes containing B56 family regulatory subunits (PP2A-B56). The conservation of Vif-mediated PP2A- B56 degradation throughout globally circulating HIV-1 subtypes suggests that it is functionally important. However, there remains a gap in understanding the mechanisms by which PP2A-B56 degradation confers a fitness advantage to HIV-1. Furthermore, in contrast to APOBEC3 degradation, the host cellular determinants required for Vif-mediated ubiquitination and degradation of PP2A-B56 are undefined. In this project, we aim to address these gaps in understanding by defining the cellular determinants and functional effects of PP2A-B56 degradation by HIV-1 Vif. In Aim 1, we will apply unbiased protein interaction technologies to determine proteins interacting with PP2A-B56 while it is degraded by Vif. In Aim 2, we will carry out a genome- wide CRISPR/Cas9 genetic screen to identify genes regulating APOBEC3 and PP2A-B56 degradation. In Aim 3, will test the impact of individual phosphorylation sites regulated by PP2A-B56 in HIV-1 replication in primary CD4+ T cells. Successful completion of this project will advance understanding of the Vif-PP2A-B56 signaling axis, potentially leading towards the development of novel classes of antiretroviral therapies that target late processes of HIV-1 infection.

项目概要 HIV-1 Vif 蛋白在感染后期表达，具有广泛描述的泛素化和降解 APOBEC3 家族中的蛋白质，从而中和其抗病毒活性。最近，另外一个我们和其他人描述了 HIV-1 Vif 的泛素化和降解蛋白磷酸酶 2A 的功能含有 B56 家族调节亚基 (PP2A-B56) 的全酶。 Vif 介导的 PP2A- 的保守 B56 在全球流行的 HIV-1 亚型中的降解表明它具有重要的功能。然而，对于 PP2A-B56 降解赋予 HIV-1 的适应性优势。此外，与 APOBEC3 降解相反，宿主细胞决定因素 Vif 介导的 PP2A-B56 泛素化和降解所需的基因尚未确定。在这个项目中，我们的目标是通过定义细胞决定因素和功能来解决这些理解上的差距。 HIV-1 Vif 对 PP2A-B56 降解的影响。在目标 1 中，我们将应用公正的蛋白质相互作用技术确定当 PP2A-B56 被 Vif 降解时与 PP2A-B56 相互作用的蛋白质。在目标 2 中，我们将进行基因组- 广泛的 CRISPR/Cas9 遗传筛选，以鉴定调节 APOBEC3 和 PP2A-B56 降解的基因。瞄准 3、将测试PP2A-B56调节的各个磷酸化位点对原代HIV-1复制的影响 CD4+ T 细胞。该项目的成功完成将促进对 Vif-PP2A-B56 信号传导的理解轴，有可能导致针对晚期的新型抗逆转录病毒疗法的开发 HIV-1 感染的过程。