ANALYSIS OF HUMAN PAPILLOMAVIRUS ENCAPSIDATION AND INFECTIVITY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 Angeleti博士的研究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)的包裹体机制。由于缺乏易于操作的系统来产生有用的病毒颗粒，对HPV感染早期步骤的遗传分析受到了阻碍。安吉莱蒂博士最近开发了一种HPV包装系统，在这种系统中，两种晚期HPV蛋白L1和L2的表达可以在酵母中产生完整的病毒粒子。该系统不仅有助于分析有效包装所需的调控元件，而且有可能用于在酵母中生产传染性HPV以供进一步研究。该项目的长期目标是了解哪些顺式和反式信号对HPV组装是重要的，以及这些信号与病毒粒子的感染性有何关系。我们假设在HPV DNA中存在一个未定义的包装元件，该元件部分或完全独立于E2。因此，我们计划：(1)确定HPV基因组中最小的顺式作用包装信号；(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E2、L1和L2对在酵母中产生的HPV假病毒感染性的影响。这些研究将提供对HPV包裹性和传染性要求的详细分析。未来的分析将允许对HPV病毒粒子的结构和功能进行详细研究，以及分析病毒在人类角质形成细胞中的附着、进入和建立。基于酵母的方法可能适用于其他病毒系统，因此，为与NCV的同事合作提供了一个很好的机会。克林顿·琼斯博士担任安吉莱蒂博士的内部导师，克雷格·迈耶斯博士(宾夕法尼亚州立大学)担任外部导师。

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 E2, L1 and L2 on infectivity of HPV pseudovirus created in yeast. These studies will provide a detailed analysis of the requirements for HPV encapsidation and infectivity. 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 is acting as an internal mentor and Dr. Craig Meyers (Penn State University) is acting as external mentor for Dr. Angeletti.