Hijacking host cellular motors for the nuclear entry of polyomaviruses

劫持宿主细胞马达使多瘤病毒进入核

• 批准号: 10650444
• 负责人: Chelsey Cierra Spriggs
• 金额: $ 24.89万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10650444
• 关键词: Advisory Committees; Award; Biochemical; Biology; Cancer Etiology; Cell Nucleus; Cells; Cellular biology; Crowding; Cytoplasm; Cytosol; DNA Tumor Viruses; DNA Viruses; Data; Disease; Dynein ATPase; Electron Microscopy; Endoplasmic Reticulum; Endosomes; Environment; Family; GAG Gene; Genetic Transcription; Genome; Genomics; Human; Infection; Intracellular Transport; Kinesin; Knowledge; Learning; Left; Life Cycle Stages; Lytic Phase; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Mentors; Merkel Cells; Merkel cell carcinoma; Michigan; Microscopy; Microtubules; Mitosis; Mitotic; Motor; Nuclear; Nuclear Envelope; Nuclear Import; Nuclear Pore Complex; Pathway interactions; Penetration; Phase; Play; Polyomavirus; Polyomavirus Infections; Process; Proteins; Public Health; Reporting; Research; Resolution; Resource Development; Role; Route; Running; Simian virus 40; Skin Carcinoma; Techniques; Time; Training; Universities; Viral; Viral Genome; Virus; Virus Replication; event cycle; human disease; human pathogen; innovation; light microscopy; metaplastic cell transformation; novel; programs; protein transport; research facility; therapeutic target; virology; virus host interaction

Project Summary/Abstract Polyomaviruses (PyVs) are small DNA tumor viruses that cause debilitating disease in humans, including the often-fatal Merkel cell carcinoma by Merkel cell polyomavirus (MCPyV). To cause infection, these non-enveloped viruses must transport through the crowded host cellular environment to reach the nucleus where transcription and replication of the viral genome leads to lytic infection or cellular transformation. During entry, PyV, is endocytosed and trafficked to the endoplasmic reticulum (ER) where it penetrates the ER membrane to reach the cytosol. Once in the cytosol, the virus is disassembled and transported to the nucleus where it is thought to enter the nucleus through the nuclear pore complex (NPC). How PyV reaches the nuclear membrane and is subsequently imported into the nucleus have yet to be determined. Intracellular transport is mediated largely through the actions of the kinesin and cytoplasmic dynein host motor proteins that transport cellular cargo towards the periphery and center of the cell, respectively. Studies of the archetype PyV, simian virus 40 (SV40), revealed that the virus exploits these motors in escaping the ER to reach the cytosol and for virus disassembly in this compartment. This K99/R00 proposal seeks to understand the role of cellular motors in the subsequent transport and nuclear entry of PyVs. During the mentored phase of this award, Dr. Spriggs will determine how kinesin-1 and dynein coordinate the transport of disassembled SV40 to the host nucleus and the mechanism by which it enters the nucleus through the NPC. At this time, she will receive training in high-resolution microscopy techniques, including correlative light and electron microscopy (CLEM), that when used in combination with both traditional and state-the-art biochemical approaches, will lead to impactful discoveries in viral and cellular nuclear import mechanisms. While much of our knowledge of human PyV infection has come from the study of SV40, key differences have been observed between it and MCPyV, the only PyV definitely associated with human cancer. One prominent distinction lies in its proposed mechanism of nuclear entry, which may instead require mitotic nuclear envelope breakdown. Despite its impact on public health, little is known of the basic biology of MCPyV infection. During the independent phase, and with her K99 training, Dr. Spriggs will delineate the entry pathway of this distinct human pathogen, culminating in a mechanistic understanding of its nuclear entry. When combined with her strong background in virology and cell biology, learning high-resolution microscopy should fully equip Dr. Spriggs to run a successful independent research program studying virus-host interactions. Further, along with her highly motivated Advisory Committee, the University of Michigan provides an ideal environment for her training, given its exceptional research facilities and professional development resources.

项目概要/摘要 多瘤病毒 (PyV) 是一种小型 DNA 肿瘤病毒，可导致人类衰弱性疾病，包括 默克尔细胞多瘤病毒 (MCPyV) 导致的通常致命的默克尔细胞癌。为了引起感染，这些无包膜的 病毒必须穿过拥挤的宿主细胞环境才能到达细胞核，在那里进行转录 病毒基因组的复制导致裂解性感染或细胞转化。在输入过程中，PyV 是 被内吞并转运至内质网 (ER)，在此穿透 ER 膜到达 细胞质。一旦进入细胞质，病毒就会被分解并运输到细胞核，并被认为在那里 通过核孔复合体（NPC）进入细胞核。 PyV 如何到达核膜 随后输入细胞核的情况尚未确定。细胞内运输很大程度上是介导的 通过运输细胞货物的驱动蛋白和细胞质动力蛋白宿主运动蛋白的作用 分别朝向细胞的外围和中心。对原型 PyV、猿猴病毒 40 (SV40) 的研究， 研究表明，病毒利用这些马达逃离内质网到达细胞质并进行病毒分解 在这个隔间里。该 K99/R00 提案旨在了解细胞马达在后续发展中的作用 PyV 的运输和入核。在该奖项的指导阶段，斯普里格斯博士将决定如何 驱动蛋白-1和动力蛋白协调分解的SV40向宿主细胞核的转运及其机制 它通过 NPC 进入细胞核。此时，她将接受高分辨率显微镜的培训 技术，包括关联光学和电子显微镜（CLEM），当与两者结合使用时 传统和最先进的生化方法，将在病毒和细胞核方面带来有影响力的发现 进口机制。虽然我们对人类 PyV 感染的大部分了解都来自对 SV40 的研究， 观察到它与 MCPyV 之间的关键差异，MCPyV 是唯一与人类明确相关的 PyV 癌症。一个显着的区别在于其提出的核进入机制，这可能需要 有丝分裂核膜破裂。尽管它对公共卫生有影响，但人们对它的基本生物学知之甚少。 MCPyV 感染。在独立阶段，通过她的 K99 培训，Spriggs 博士将描绘进入 这种独特的人类病原体的途径，最终对其进入核的机制有了了解。什么时候 结合她在病毒学和细胞生物学方面的深厚背景，学习高分辨率显微镜应该 斯普里格斯博士完全有能力开展一项成功的独立研究项目，研究病毒与宿主的相互作用。 此外，连同她积极主动的咨询委员会，密歇根大学提供了一个理想的 鉴于其卓越的研究设施和专业发展资源，为她的培训提供了良好的环境。