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。更 迄今已确定了150多个类型。根据人口和年龄， 高达20%的女性和60%的男性可以携带一种或多种HPV 类型HPV感染上呼吸道和肛门生殖器粘膜组织 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如16型中，两种功能蛋白， E6和E7协同作用，使细胞永生， 表型转化受感染的宿主细胞。然而，E6是 推动这些变化的主要因素。有HPV 16基因组变体， 值得注意的是，E6基因比E7基因具有更多的多态性。HPV变异体已被 与它们首次被发现的地理区域有关，这表明， 它们与特定的人类群体共同进化。我们假设这些 E6变体影响相应变体E6的性能 蛋白质，这很可能是由于蛋白质的构象变化， 结构事实上，我们最近的功能研究表明， 转化潜力和能力，坚持在主机所造成的只是3 2种HPV 16变异体中E6蛋白N-末端的氨基酸变化：欧洲 原型（EP）和亚裔美国人（AA），这两种人在美国人群中很常见。我们的目标是这个跨学科的 研究计划，在三个短期目标解决，是为了了解 基因组HPV变异的潜在机制，重点是E6， 病毒-宿主相互作用的显著变化，影响其 转化受感染的宿主细胞我们将使用几种体外细胞培养模型 （用单个E6变体基因和3D转染的单层细胞 用全长HPV 16变异基因组中的任一种转染的筏培养物） 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。的 这项研究计划的长期目标将是：1）促进知识， 基础病毒研究和生物信息学，重点是进化和 转化DNA病毒的分子机制，2）提供定制 “生物学家友好”的生物信息学软件用于病毒宿主分析和3）提供多才多艺的高素质人才（HQP）谁拥有的社会，智力和实践技能，需要在今天的快节奏的科学界取得成功。我们的成功将是 在我们强大的跨学科研究环境的保证 研究所（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），对所提出的工作具有专业知识，出色的HQP监督，最先进的 设备以及国家和国际合作者。

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.