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.

项目摘要/摘要 多瘤病毒（PYVS）是小的DNA肿瘤病毒，引起人类衰弱的疾病，包括 默克尔细胞多瘤病毒（MCPYV）通常致命的默克尔细胞癌。为了引起感染，这些不发达 病毒必须通过拥挤的宿主细胞环境传输才能到达转录的细胞核 病毒基因组的复制导致裂解感染或细胞转化。进入期间，PYV是 内吞和贩运到内质网（ER），在那里穿透ER膜即可到达 细胞质。一旦进入细胞质，病毒就被拆卸并转运到被认为是的细胞核 通过核孔复合物（NPC）进入细胞核。 PYV如何到达核膜，是 随后进口到核尚未确定。细胞内转运在很大程度上介导 通过驱动蛋白和细胞质动力蛋白的作用，可传输细胞货物 分别朝向细胞的外围和中心。研究原型PYV的研究，Simian病毒40（SV40）， 揭示该病毒利用这些电动机逃脱了ER到达细胞质和病毒拆卸 在这个隔间中。该K99/R00提案旨在了解蜂窝电机在随后的 PYVS的运输和核进入。在该奖项的指导阶段，Spriggs博士将确定如何 驱动蛋白1和动力蛋白协调拆卸的SV40向宿主核的运输，并通过 它通过NPC进入核。目前，她将接受高分辨率显微镜的培训 技术，包括相关光和电子显微镜（CLEM），当将两者结合使用时 传统和国家的生化方法将导致病毒和细胞核的有影响力的发现 进口机制。尽管我们对人类PYV感染的许多了解来自对SV40的研究，但 已经观察到它与MCPYV之间观察到的主要差异，这是唯一与人类相关的PYV 癌症。一个明显的区别在于其拟议的核进入机制，这可能需要 有丝分裂核包膜崩溃。尽管它对公共卫生有影响，但对 MCPYV感染。在独立阶段，在K99培训的情况下，Spriggs博士将描述该条目 这种独特的人类病原体的途径，最终导致对其核进入的机械理解。什么时候 结合她在病毒学和细胞生物学方面的良好背景，学习高分辨率显微镜应该 Spriggs博士完全装备了一项成功研究病毒宿主相互作用的独立研究计划。 此外，密歇根大学与她积极进取的咨询委员会一起提供了理想 鉴于其卓越的研究设施和专业发展资源，她的培训环境。