Characterization of the antiviral and nuclear regulatory functions of FEZ1 & NEK1

描述（由申请人提供）：人类免疫缺陷病毒1型（HIV-1）是逆转录病毒科的成员，可引起获得性免疫缺陷综合征（AIDS），影响全球3330万人。像许多病毒一样，逆转录病毒需要进入宿主细胞核才能复制，但对所涉及的过程和因素仍知之甚少。病毒颗粒进入宿主细胞后，通过细胞质进入细胞核 微管是围绕位于细胞核附近的微管组织中心（MTOC）排列的长丝状运输网络。我们最近的工作已经确定了一种神经元蛋白FEZ 1，它定位于MTOC并通过阻止病毒DNA进入细胞核来抑制包括HIV-1在内的许多逆转录病毒的感染。我们的初步数据还表明，FEZ 1相互作用蛋白，NEK 1具有类似的神经元表达模式和对HIV-1感染的抑制作用，这表明FEZ 1和至少一些其相互作用蛋白可能形成神经元复合物的一部分，损害逆转录病毒感染。我们额外的初步数据显示，FEZ 1还抑制另一种临床上重要的嗜神经DNA病毒，单纯疱疹病毒1型（HSV-1）的感染，这表明FEZ 1在神经元中的表达可能限制了它们对许多不同病毒感染的易感性。最后，FEZ 1在小胶质细胞（另一种脑细胞类型）中的异位表达会导致核结构的变化，称为多小叶，这可能有助于其抗病毒特性，并表明FEZ 1及其相互作用蛋白可能调节重要的，但对核本身的功能知之甚少。在这项提案中，我们的目标是表征抗病毒和核 FEZ 1的调控功能，详细定义参与调控感染的结构域以及参与多小叶核形成的结构域，并确定宿主信号传导途径对FEZ 1功能的调控。通过研究其他FEZ 1和NEK 1相互作用因子对所观察到的表型的贡献，我们希望构建这些蛋白质如何调节病毒感染和核结构或运动的图像，这将为与MTOC相关的因子如何与病毒感染和大型货物（如病毒衣壳）的更广泛运动重要的核功能进行通信和调节提供重要的见解，进入细胞核。因此，这些研究也可能有助于我们更广泛地了解核运输和结构的控制。

DESCRIPTION (provided by applicant): Human immunodeficiency virus type 1 (HIV-1) is a member of the retroviridae that causes Acquired Immunodeficiency Syndome (AIDS), which affects over 33.3 million people worldwide. Like many viruses, retroviruses require access to the host nucleus in order to replicate yet the processes and factors involved remain poorly understood. After entry into the cell, viral particles transit the cytoplasm to the nucleus on host microtubules, long filamentous transport networks that arrange around the microtubule organizing centre (MTOC) located near the nucleus. Our recent work has identified a neuronal protein, FEZ1 that localizes to the MTOC and suppress infection by a number of retroviruses, including HIV-1, by blocking the entry of viral DNA into the nucleus. Our preliminary data also suggests that the FEZ1 interacting protein, NEK1 has a similar neuronal expression pattern and inhibitory effect on HIV-1 infection, suggesting that FEZ1 and at least some of its interacting proteins may form part of a neuronal complex that impairs retroviral infection. Our additional preliminary data shows that FEZ1 also suppresses infection by another clinically important neurotropic DNA virus, Herpes Simplex Virus type 1 (HSV-1), suggesting that FEZ1 expression in neurons may limit their susceptibility to infection by a number of distinct viruses. Finally, ectopic expression of FEZ1 in microglia, another brain cell type, causes changes in nuclear architecture, called multi-lobulation, which may contribute to its antiviral properties and which suggests that FEZ1 and its interacting proteins may regulate important, poorly understood functions of the nucleus itself. In this proposal, we aim to characterize the antiviral and nuclear regulatory functions of FEZ1 in detail, defining the domains involved in regulating infection as well as those involved in the formation of multi-lobulated nuclei, and determining the regulation of FEZ1 function by host signaling pathways. By also examining the contribution of additional FEZ1- and NEK1-interacting factors to the phenotypes observed, we hope to build a picture of how these proteins regulate viral infection and nuclear architecture or movement, which will provide important insights into how factors associated with the MTOC communicate with and regulate nuclear functions important for viral infection and the broader movement of large cargoes, such as viral capsids, into the nucleus. As such, these studies are also likely to contribute to our broader understanding of the control of nuclear transport and architecture.