The role of FEZ1 in early HIV-1 infection

FEZ1 在早期 HIV-1 感染中的作用

基本信息

批准号：
10647657
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
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-24 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10647657
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 Resource-limited setting Reverse Transcription Role T-Lymphocyte Testing Universities Viral Viral Genome Work antiretroviral therapy cell motility cell type cofactor design genetic regulatory protein innovation insight mutant novel strategies overexpression particle preference recruit

项目摘要

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）的广泛使用有效地增加了许多受感染的人HIV-1在发达和贫困中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开发新方法治疗感染。 HIV-1员工异常，复杂地交织的早期感染涉及逆转录的策略，Capsid Core（也称为“无涂层”）和尽管其确切的时机和位置仍然有争议，但越来越多的证据表明至少部分脱膜发生在传输到核过程中的细胞质中。的确，传入的HIV-1颗粒暴露于微管（MT）的双向运动暗示其关联随着内（动力蛋白）和外部（动力蛋白）MT电动机，最近的研究表明对立这些电动机产生的力有助于解涂。尽管如此，HIV-1似乎没有绑定电动机直接但使用的是，直到近年来，其身份一直保持着神秘的方式。我们的工作在上一个周期中资助将HIV-1驱动蛋白-1适配器确定为fasiculation and伸长因子zeta 1（fez1）。我们进一步确立了FEZ1在传入病毒颗粒的运输和脱覆膜中的核心作用在自然靶细胞中，通过控制驱动蛋白-1活性的FEZ1磷酸化受调节。此外，我们发现HIV-1核与微管相关的调节激酶2（MARK2）与局部控制病毒颗粒上的FEZ1磷酸化。我们进一步表明，HIV-1颗粒也高度结合专门的MT调节蛋白诱导稳定MT网络的形成，这是MT细丝的子集受到动机电动机的青睐。使用创新的结构和功能研究与Xiong合作耶鲁大学的实验室，我们的初步数据揭示了HIV-1使用的常见且高亲和力的约束策略参与FEZ1进行运输，该运输是由Capsid Hexamers介导的，是FEZ1的四个盘绕线圈域之一。数据还表明，Fez1和Mark2以控制FEZ1的程度的方式竞争结合 HIV-1衣壳上的磷酸化。此外，我们确定了一个新的宿主因素，我们的数据表明绑定了 Fez1中不同的盘绕螺旋区域，并通过传入的病毒颗粒来利用以增强MT稳定细胞周围。累积地，我们的数据表明，Fez1中不同的盘绕螺旋域Mediate Capsid 结合，运动募集和MT稳定，以协调早期HIV-1运输的几个方面脱涂。在此提案中，我们旨在确定Fez1和Mark2在HIV-1上的作用如何促进早期感染并扩展我们的新发现，即FEZ1在早期起着多功能作用通过募集MT稳定性的电动机和调节剂来感染传入的HIV-1颗粒。结果我们的研究将为FEZ1的多功能性提供重要的机械见解并扩展我们对HIV-1如何控制天然靶细胞的早期感染中如何控制几个重要步骤。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Microtubule plus end-associated CLIP-170 initiates HSV-1 retrograde transport in primary human cells.

DOI：
10.1083/jcb.201505123
发表时间：
2015-10-26
期刊：
The Journal of cell biology
影响因子：
0
作者：
Jovasevic V;Naghavi MH;Walsh D
通讯作者：
Walsh D

Exploitation of Cytoskeletal Networks during Early Viral Infection.

早期病毒感染期间细胞骨架网络的利用。

DOI：
10.1016/j.tim.2018.06.008
发表时间：
2019-01
期刊：
Trends in microbiology
影响因子：
15.9
作者：
Walsh D;Naghavi MH
通讯作者：
Naghavi MH

Dynactin 1 negatively regulates HIV-1 infection by sequestering the host cofactor CLIP170.

Dynactin 1 通过隔离宿主辅因子 CLIP170 来负向调节 HIV-1 感染。

DOI：
10.1073/pnas.2102884118
发表时间：
2021
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Shanmugapriya,Shanmugapriya;SantosdaSilva,Eveline;Campbell,JacksonA;Boisjoli,Marie-Philipe;Naghavi,MojganH
通讯作者：
Naghavi,MojganH

HIV-1 capsid exploitation of the host microtubule cytoskeleton during early infection.

DOI：
10.1186/s12977-021-00563-3
发表时间：
2021-07-06
期刊：
Retrovirology
影响因子：
3.3
作者：
Naghavi MH
通讯作者：
Naghavi MH

HIV-1 induces the formation of stable microtubules to enhance early infection.

DOI：
10.1016/j.chom.2013.10.012
发表时间：
2013-11-13
期刊：
Cell host & microbe
影响因子：
30.3
作者：
Sabo Y;Walsh D;Barry DS;Tinaztepe S;de Los Santos K;Goff SP;Gundersen GG;Naghavi MH
通讯作者：
Naghavi MH