ANALYSIS OF HUMAN PAPILLOMAVIRUS ENCAPSIDATION AND INFECTIVITY

人乳头瘤病毒衣壳和感染性分析

• 批准号: 7381208
• 负责人: Peter C. Angeletti
• 金额: $ 15.57万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381208
• 关键词: ANALYSIS; HUMAN; PAPILLOMAVIRUS; ENCAPSIDATION; INFECTIVITY

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Dr. Angeletti?s research investigates the mechanism of encapsidation of the persistent infecting virus, human papillomavirus (HPV) by utilizing a novel yeast-based system. The genetic analysis of early steps in HPV infection has been hampered by the lack of easily manipulatable systems in which to generate useful amounts of viral particles. Dr. Angeletti has recently developed an HPV-packaging system in which expression of two late HPV proteins, L1 and L2 proteins can generate intact virions in yeast. This system is not only useful for analysis of regulatory elements required for efficient packaging, but potentially in production of infectious HPV in yeast for further study. The long-term goal of this project is to understand what cis and trans acting signals are important for HPV assembly and how these relate to infectivity of virions. We hypothesize that an undefined packaging element exists in HPV DNA that is either partially or entirely independent of E2. Therefore, we plan to: (1) define the minimal cis-acting packaging signal in the HPV genome, (2) determine the role of trans-acting factors E2 and L2 in efficient DNA packaging, (3) determine the effect of L1 and L2 genotype-specific differences on infectivity of HPV pseudovirus created in yeast. Different HPV genotypes; lab strains and natural isolates derived from patient samples from Zambia Africa will be used as a source of capsid genes. These studies will provide a detailed analysis of the requirements for HPV encapsidation and infectivity that can be related to the known pathogenicity of HPV strains. Future analyses will allow detailed structure-function studies of HPV virions as well as analysis of viral attachment, entry and establishment in human keratinocytes. The yeast-based approach is potentially adaptable to other viral systems and therefore, provides a great opportunity for collaboration with NCV colleagues. Dr. Clinton Jones will act as a mentor for Dr. Angeletti. Dr. Jones has a breadth of experience in studies of persistent DNA viruses and will provide advice on the strategies employed in these studies.??

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。安杰莱蒂医生的研究通过利用一种新的基于酵母的系统来研究持续感染病毒人乳头瘤病毒（HPV）的灭活机制。由于缺乏易于操作的系统来产生有用数量的病毒颗粒，HPV感染早期步骤的遗传分析受到阻碍。Angeletti博士最近开发了一种HPV包装系统，其中两种晚期HPV蛋白L1和L2蛋白的表达可以在酵母中产生完整的病毒体。该系统不仅可用于分析有效包装所需的调控元件，而且可能用于进一步研究酵母中感染性HPV的生产。该项目的长期目标是了解哪些顺式和反式作用信号对HPV组装很重要，以及这些信号如何与病毒体的感染性相关。我们假设HPV DNA中存在一个不确定的包装元件，它部分或完全独立于E2。因此，我们计划：（1）确定HPV基因组中最小顺式作用包装信号，（2）确定反式作用因子E2和L2在有效DNA包装中的作用，（3）确定L1和L2基因型特异性差异对酵母中产生的HPV假病毒感染性的影响。不同的HPV基因型;来自非洲赞比亚患者样本的实验室菌株和天然分离株将用作衣壳基因的来源。这些研究将提供HPV感染和感染性要求的详细分析，这些要求可能与HPV毒株的已知致病性有关。未来的分析将允许HPV病毒体的详细结构-功能研究以及病毒在人角质形成细胞中的附着、进入和建立的分析。基于酵母的方法可能适用于其他病毒系统，因此，为与NCV同事的合作提供了很好的机会。克林顿·琼斯博士将担任安杰莱蒂博士的导师。琼斯博士在持久性DNA病毒的研究方面拥有广泛的经验，并将为这些研究中采用的策略提供建议。