The role of FEZ1 in early HIV-1 infection

FEZ1 在早期 HIV-1 感染中的作用

• 批准号: 10207439
• 负责人: Mojgan Hosseini Naghavi
• 金额: $ 36.02万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-24 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207439
• 关键词: Adaptor Signaling Protein; Affect; Affinity; Binding; Binding Proteins; C-terminal; Capsid; Cell Line; Cell Nucleus; Cells; Coiled-Coil Domain; Collaborations; Competitive Binding; Complex; Coupled; Cytoplasm; Data; Development; Dynein ATPase; Elongation Factor; Equilibrium; Event; Exhibits; Filament; Funding; HIV Infections; HIV-1; Human; Image; In Vitro; Individual; Infection; Integration Host Factors; Kinesin; Location; Longevity; Mediating; Microglia; Microtubule Stabilization; Microtubules; Modification; Motor; Nature; Nucleocapsid; Outcome; Phosphorylation; Phosphotransferases; Play; Plus End of the Microtubule; Process; Proteins; Public Health; Resources; Reverse Transcription; Role; Structure; Suggestion; T-Lymphocyte; Testing; Universities; Viral; Viral Genome; Work; antiretroviral therapy; base; cell motility; cell type; cofactor; design; genetic regulatory protein; innovation; insight; mutant; novel strategies; overexpression; particle; preference; recruit

Although widespread use of combination antiretroviral therapy (cART) has effectively increased the life span of many infected individuals, HIV-1 continues to be a major public health issue in both developed and poor resource settings. As such, understanding the basic mechanisms of its replication cycle is instrumental to the development of new approaches to treat infection. HIV-1 employs unusual, intricately intertwined early infection strategies involving reverse transcription, disassembly of capsid core (also known as “uncoating”) and transport to the nucleus. Although its precise timing and location remain contentious, growing evidence suggests that at least partial uncoating occurs in the cytoplasm during transport to the nucleus. Indeed, incoming HIV-1 particles exhibit microtubule (MT) based bi-directional motility suggestive of their association with both inward (dynein) and outward (kinesin) MT motors, and recent studies suggest that the opposing forces generated by these motors facilitate uncoating. Despite this, HIV-1 does not appear to bind motors directly but instead, uses motor adaptors whose identity remained enigmatic until recent years. Our work funded in the previous cycle identified the HIV-1 kinesin-1 adaptor as Fasiculation and Elongation Factor Zeta 1 (FEZ1). We further established FEZ1’s central role in the transport and uncoating of incoming viral particles in natural target cells, which is regulated through FEZ1 phosphorylation that controls kinesin-1 activity. Moreover, we found that HIV-1 cores bind microtubule associated regulatory kinase 2 (MARK2) to locally control FEZ1 phosphorylation on viral particles. We further showed that HIV-1 particles also bind highly specialized MT regulatory proteins to induce the formation of stable MT networks, a subset of MT filaments favored by kinesin motors. Using innovative structural and functional studies in collaboration with the Xiong Lab at Yale University, our preliminary data reveals an usual and high affinity binding strategy used by HIV-1 to engage FEZ1 for transport that is mediated by capsid hexamers and one of four coiled-coil domains in FEZ1. Data also suggests that FEZ1 and MARK2 compete for binding in a manner that controls the extent of FEZ1 phosphorylation on HIV-1 capsids. In addition, we identify a new host factor that our data suggests binds distinct coiled-coil regions in FEZ1 and is exploited by incoming viral particles to enhance MT stabilization at the cell periphery. Cumulatively, our data suggests that distinct coiled-coil domains in FEZ1 mediate capsid binding, motor recruitment and MT stabilization to coordinate several aspects of early HIV-1 transport and uncoating. In this proposal, we aim to determine how FEZ1 and MARK2 function on the HIV-1 capsid to promote early infection and expand upon our new findings that FEZ1 plays a multifunctional role in early infection by recruiting both motors and regulators of MT stability to incoming HIV-1 particles. The outcome of our studies will provide important mechanistic insights into the multifunctionality of FEZ1 and expand our broader understanding of how HIV-1 controls several important steps in early infection of natural target cells.

尽管联合抗逆转录病毒疗法(CART)的广泛使用有效地延长了患者的寿命 在许多感染者中，艾滋病毒-1在发达国家和穷国仍然是一个主要的公共卫生问题 资源设置。因此，了解其复制周期的基本机制对于 开发治疗感染的新方法。HIV-1使用不寻常的、错综复杂的早期感染 涉及反转录、衣壳核心的拆解(也称为“脱壳”)和 运送到原子核。尽管它的确切时间和地点仍然存在争议，但越来越多的证据表明 这表明，在运输到细胞核的过程中，细胞质中至少发生了部分脱涂层。的确， 进入的HIV-1颗粒表现出基于微管(MT)的双向运动，提示它们之间存在关联 对于内向(动力蛋白)和向外(动力蛋白)的MT马达，最近的研究表明，相反的 这些马达产生的力有助于脱膜。尽管如此，HIV-1似乎并没有绑定马达 直接，但取而代之的是使用电机适配器，直到最近几年，这种适配器的身份一直是个谜。我们的工作 在上一个周期资助的确定HIV-1激动素-1接头为变形和延伸因子Zeta 1(FEZ1)。我们进一步确立了FEZ1的S在传入病毒颗粒的运输和剥离中的核心作用 在天然靶细胞中，这是通过FEZ1磷酸化来调节的，该磷酸化控制着kinesin-1的活性。 此外，我们还发现HIV-1核心与微管相关调节蛋白2(Mark2)结合在一起 控制病毒颗粒上FEZ1的磷酸化。我们进一步表明，HIV-1颗粒也高度结合 专门的MT调节蛋白诱导形成稳定的MT网络，MT细丝的一个子集 受到动蛋白马达的青睐。与熊学院合作进行创新的结构和功能研究 我们的初步数据揭示了HIV-1使用的一种常见的高亲和力结合策略来 与FEZ1接合，进行由衣壳六聚体和FEZ1中四个卷曲结构域之一介导的转运。 数据还表明，FEZ1和Mark2以控制FEZ1范围的方式竞争结合 HIV-1衣壳上的磷酸化。此外，我们确定了一个新的宿主因素，我们的数据表明该因素与 FEZ1中独特的螺旋卷曲区域，并被传入的病毒颗粒利用来增强MT的稳定性 细胞的外围。总体而言，我们的数据表明，FEZ1中不同的螺旋线圈结构域参与衣壳的调节 结合、运动招募和MT稳定以协调HIV-1早期运输和 揭开涂层。在这项提案中，我们的目标是确定FEZ1和Mark2如何在HIV-1衣壳上发挥作用 促进早期感染并扩展我们的新发现，即FEZ1在早期感染中发挥多功能作用 通过招募发动机和MT稳定性调节器对传入的HIV-1颗粒进行感染。其结果是 我们的研究将为FEZ1的多功能提供重要的机制见解，并扩展我们的 更广泛地了解HIV-1如何控制自然靶细胞早期感染的几个重要步骤。