Virus-host interactions in the context of natural human papillomavirus type 16 E6 gene variants

人乳头瘤病毒(HPV)是影响人类的最常见微生物感染之一。*到目前为止，已鉴定出150多种类型。根据人口和年龄的不同，*多达20%的女性和60%的男性可能携带一种或多种HPV*型。HPV感染上呼吸道和肛门生殖道的粘膜组织以及皮肤。它们的基因组编码几种结构和功能蛋白质。在某些HPV中，如16型，两种功能蛋白*E6和E7协同作用，使受感染的宿主细胞永生和*表型转化。然而，E6是推动这些变化的主要影响因素。有HPV16基因组变种，值得注意的是，E6基因比E7基因更具多态。人类乳头状瘤病毒的变种被发现与最初发现它们的地理区域有关，这表明它们与特定的人类群体共同进化。我们假设这些*E6变异体影响相应的变异体E6*蛋白的性能，这很可能是由于蛋白质*结构的构象变化。事实上，我们最新的功能研究表明，仅由两个HPV16变种中E6蛋白N端的3*氨基酸变化引起的不同*转化潜力和在宿主中持续存在的能力：欧洲*原型(EP)和亚裔美国人(AA)，这两种变体都经常在美国人中发现。我们这项跨学科的研究计划有三个短期目标，我们的目标是了解基因组HPV变异如何引起病毒-宿主相互作用的重大变化的潜在机制，这些变异主要集中在E6上，这些变化影响了它们转化受感染宿主细胞的能力。我们将使用几种体外细胞培养模型*(转导单个E6变异基因的单层细胞和转染任一全长HPV16变异基因组的3D*RAFT培养)*和方法学方法(高通量基因表达分析，包括*RNA测序、生物信息学、代谢途径表征和*蛋白质-蛋白质相互作用调查)来回答我们的研究问题。该研究计划的长期目标将是：1)增进基础病毒研究和生物信息学方面的知识，重点是转化DNA病毒的进化和分子机制；2)提供用于病毒宿主分析的定制的“生物学家友好的”生物信息学软件；3)提供具备在当今快节奏的科学界取得成功所需的社交、智力和动手技能的多才多艺的高素质人员(HQP)。我们的成功将得到我们研究所强大的跨学科研究环境(分子生物学和病毒学、细胞生物学、生物化学、生物信息学和*组织学)的*保障。建议工作的专业知识、出色的HQP监督、最先进的设备以及国内和国际合作者。**

Human papillomavirus*(HPV) is one of the most common microbial infections affecting humans. More*than 150 types have been identified to date. Depending on population and age,*up to 20% of females and 60% of males can harbour one or more HPV*types. HPVs infect mucosal tissues of the upper respiratory and anogenital*tracts as well as the skin. Their genomes encode several structural and*functional proteins. In certain HPVs such as type 16, two functional proteins,*E6 and E7, act synergistically to immortalize and and*phenotypically transform infected host cells. However, E6 is*the main effecter driving these changes. There are HPV16 genome variants and,*notably, the E6 gene is more polymorph than the E7 gene. HPV variants have been*linked to the geographical region where they were first found, suggesting that*they have co-evolved with specific human populations. We hypothesize that these*E6 variants influence the performance of the corresponding variant E6*proteins, which is most likely due to conformational changes in protein*structure. Indeed, our most recent functional studies suggest differential*transforming potential and ability to persist in the host caused by just 3*amino acid changes in the N-terminus of the E6 protein in 2 HPV16 variants: European*Prototype (EP) and Asian American (AA), both of which are frequently found in American populations. Our goal with this inter-disciplinary*research program, addressed in three short-term objectives, is to understand*the underlying mechanisms of how genomic HPV variants, with focus on E6, cause*significant changes in virus-host interactions which affect their ability to*transform infected host cells. We will use several in vitro cell culture models*(monolayer cells transduced with single E6 variant genes and 3D*raft cultures transfected with either of the full-length HPV16 variant genomes)*and methodological approaches (high-throughput gene expression analyses including*RNA-sequencing, bioinformatics, metabolic pathway characterization and*protein-protein interaction surveys) to answer our research questions. The*long-term objectives of this research program will be to 1) advance knowledge in*basic virus research and bioinformatics, with a focus on the evolution and*molecular mechanisms of transforming DNA viruses, 2) offer custom*"biologist-friendly" bioinformatics software for virus-host analyses and 3) provide versatile highly qualified personnel (HQP) who have the social, intellectual and hands-on skills required to be successful in today's fast paced scientific community. Our success will be*guaranteed by the strong inter-disciplinary research environment at our*institute (molecular biology and virology, cell biology, biochemistry, bioinformatics and*histology), expertise in the work proposed, excellent HQP supervision, state-of-the-art*equipment as well as national and international collaborators. **