Mechanisms and Consequences of L2-Dependent Subcellular Trafficking of the HPV Genome.

HPV 基因组 L2 依赖性亚细胞贩运的机制和后果。

• 批准号: 10613448
• 负责人: Samuel K Campos
• 金额: $ 33.09万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10613448
• 关键词: Address; Adopted; Anogenital cancer; Bacterial Toxins; Bacterial Translocation; Binding; Biological; Biology; Capsid; Cell Nucleus; Cell Proliferation; Cell surface; Cells; Cellular Membrane; Cellular biology; Centrioles; Centrosome; Chromatin; Chromosome Condensation; Chromosomes; Complex; Cutaneous; Cytosol; DNA; DNA Viruses; Data; Detection; Double Stranded DNA Virus; Elements; Endosomes; Ensure; Epithelium; Etiology; Event; Family; Foundations; Future; Genetic Materials; Genome; Goals; Golgi Apparatus; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus HPV L1 protein; Hydrophobicity; Immune; Immunologic Surveillance; Infection; Innate Immune Response; Interferons; Interphase; Knowledge; L2 viral capsid protein; Life Cycle Stages; Locales; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of nasopharynx; Mediating; Membrane; Membrane Proteins; Metaphase; Microtubules; Minor; Mitosis; Mitotic; Mitotic Chromosome; Modeling; Molecular; Molecular Conformation; Mucous Membrane; N-terminal; Nature; Nuclear Envelope; Oncogenic; Pathway interactions; Penetration; Peptide Hydrolases; Process; Prometaphase; Proteolysis; Role; Sensory; Sexually Transmitted Diseases; Site; Sorting; Stimulator of Interferon Genes; Structure; System; Time; Tissues; Transmembrane Domain; United States; Vaccination; Vesicle; Viral; Virion; Virus; Woman; Work; chronic infection; daughter cell; dimer; ds-DNA; fascinate; high risk; innate immune pathways; innovation; insight; keratinocyte; keratinocyte differentiation; men; migration; novel; pathogen; recruit; residence; response; sensory system; sorting nexins; trafficking; trans-Golgi Network

Human papillomaviruses (HPVs) are the most common sexually transmitted infection. These viruses infect and replicate in mucosal and cutaneous epithelium, inducing cell proliferation as part of their replicative life cycle. High risk oncogenic HPVs caused over 500,000 cases of cervical cancer worldwide, the fourth most common cancer in women (2012 estimates). HPVs are small DNA viruses and must deliver their genomes to the host cell nucleus to initiate a successful infection. Like all other non-enveloped viruses, HPVs must transport their genetic material (vDNA) across an intracellular limiting membrane, a critical event mediated by the minor capsid protein L2. Our prior work has implicated the N-terminal domain of L2 as a crucial region for this membrane penetration activity. N- terminal cleavage of L2 by the host cell protease furin and a conserved transmembrane domain (TMD) are essential for vDNA translocation. During virion entry, cleavage of L2 by cell surface furin appears to modulate a conformational change enabling L2 to interact and span across local membranes, an activity we call “protrusion”. In this membrane-protruding conformation, L2 is able to access the cytosol to recruit cellular sorting factors while remaining complexed to the lumenal vDNA. Engagement of cytosolic sorting nexins and retromer enable efficient L2-dependent transport of vDNA to the Golgi. Our recent work reveals that this membrane protruding L2/vDNA complex remains at the Golgi until the onset of mitosis, when the Golgi naturally disperses coincident with chromosome condensation, centriole migration, spindle assembly, and nuclear envelope breakdown. During this brief period of dynamic changes, the vesicle-bound membrane-protruding L2/lumenal vDNA complex egresses from the vesiculating Golgi, localizes near centrosomes by prometaphase, and appears to traverse along the spindle, reaching the condensed chromosomes by metaphase. At this time, we believe L2 uses a chromatin- binding domain to physically tether to host chromosomes, a function that is essential for full translocation of the vDNA out of the post-Golgi vesicle. This chromosome-tethered L2/vDNA then partitions into daughter cells to establish a persistent infection of basal keratinocytes. Herein, we propose studies aimed at understanding the fascinating biological mechanisms and implications of L2 subcellular trafficking and mitosis-dependent translocation. Specifically, we aim to understand the nature of the membrane-protruding L2/vDNA complex and the role of furin in achieving this conformation, as well as determine the timing and locale of mitosis- dependent translocation of L2/vDNA from post-Golgi vesicle to metaphase chromosome. Finally, we will explore the biological consequences of this unique trafficking and translocation with regards to innate immune surveillance. These fundamental processes likely contribute towards viral evasion of early innate immune responses and may influence the establishment of persistent infections- hallmarks of oncogenic HPVs that undoubtedly contributes to the oncogenic nature of these viruses.

人乳头瘤病毒（HPV）是最常见的性传播感染。这些病毒在粘膜和皮肤上皮中感染并复制，作为其复制生命周期的一部分诱导细胞增殖。高风险致癌性HPV在全球造成了超过500,000例宫颈癌，这是女性第四大癌症（2012年估计）。 HPV是小的DNA病毒，必须将其基因组传递到宿主细胞核才能成功感染。与所有其他非发育病毒一样，HPV必须将其遗传物质（VDNA）转移到细胞内极限膜上，这是由次要衣壳蛋白L2介导的关键事件。我们先前的工作已实施了L2的N末端结构域，作为该膜穿透活性的关键区域。宿主细胞蛋白素和配置的跨膜结构域（TMD）对L2的N末端切割对于VDNA易位至关重要。在进入病毒素的过程中，通过细胞表面脂蛋白对L2进行裂解似乎可以调节会议变化，从而使L2在局部膜上相互作用并跨越跨度，我们称之为“突出”。在这次膜预欺骗会议中，L2能够进入细胞质以募集细胞分选因子，同时保持与Lumenal VDNA复合。细胞溶胶分选Nexins和Demomer的参与使VDNA向高尔基体有效地依赖L2依赖性转运。我们最近的工作表明，这种突出的L2/vDNA复合物的膜保持在高尔基体，直到有丝分裂发作，当时高尔基人自然地分散了与染色体凝结，中心迁移，纺锤体组装和核包膜分解相吻合。在这一短暂的动态变化期间，囊泡结合的膜形成的L2/Lumenal VDNA复合物从囊泡高尔基体中排出，通过Prometathase将其定位在中心体附近，并且似乎沿主轴横穿，并通过转子酶达到凝结体。目前，我们认为L2使用染色质结合结构域进行物理系绳以宿主染色体，该功能对于从后高尔基体囊泡中完全翻译的功能至关重要。然后，这种染色体螺旋体的L2/vDNA分隔到子细胞中，以建立碱性角质形成细胞的持续感染。本文中，我们提出的研究旨在了解L2亚细胞运输和有丝分裂依赖性易位的迷人生物学机制和含义。具体而言，我们旨在了解膜构抑制L2/vDNA复合物的性质，以及Furin在达到这种构象中的作用，并确定L2/VDNA从后高尔基蔬菜对中期染色体的L2/VDNA的时机和位置。最后，我们将探讨这种独特的贩运和易位的生物学后果，并考虑了先天免疫监视。这些基本过程可能有助于早期先天免疫调查的病毒进化，并可能影响建立持续的感染 - 无疑有助于这些病毒的致癌性质的致癌HPV的标志。

项目成果

Master mitotic kinases regulate viral genome delivery during papillomavirus cell entry.

• DOI: 10.1038/s41467-023-35874-w
• 发表时间: 2023-01-23
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Rizzato, Matteo;Mao, Fuxiang;Chardon, Florian;Lai, Kun-Yi;Villalonga-Planells, Ruth;Drexler, Hannes C. A.;Pesenti, Marion E. E.;Fiskin, Mert;Roos, Nora;King, Kelly M. M.;Li, Shuaizhi;Gamez, Eduardo R. R.;Greune, Lilo;Dersch, Petra;Simon, Claudia;Masson, Murielle;Van Doorslaer, Koenraad;Campos, Samuel K. K.;Schelhaas, Mario
• 通讯作者: Schelhaas, Mario