Cell surface events of human papillomavirus type 16 and 18 infection

人乳头瘤病毒 16 型和 18 型感染的细胞表面事件

基本信息

批准号：
7895816
负责人：
Martin Sapp
金额：
$ 36.26万
依托单位：
LOUISIANA STATE UNIV HSC SHREVEPORT
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-17 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7895816
关键词：
Accounting Address Affect Amino Acids Antibodies Binding Binding Sites Biochemical Biological Biological Assay Biology Capsid Capsid Proteins Cell Culture Techniques Cell surface Cell-Matrix Junction Cells Cervix carcinoma Clathrin Cleaved cell Collaborations Complement Confocal Microscopy Crystallization Cyclophilins DNA Viruses Data Development Endocytosis Endosomes Epidermis Event Extracellular Matrix Family Generations Genetic Transcription Genome Goals Heparan Sulfate Proteoglycan Heparin Heparin Binding Human Papilloma Virus-Related Malignant Neoplasm Human Papillomavirus Human papilloma virus infection Human papillomavirus 16 Human papillomavirus 18 In Vitro Incidence Infection Intracellular Transport Investigation Knock-out L1 viral capsid protein L2 viral capsid protein Lesion Life Cycle Stages Los Angeles Lysine Malignant - descriptor Malignant Neoplasms Maps Mediating Medical Microtubules Minor Molecular Molecular Conformation Monoclonal Antibodies Mutagenesis N-terminal Nuclear Peptidylprolyl Isomerase Pharmaceutical Preparations Pharmacotherapy Population Process Proteins Research Roentgen Rays Role Screening for cancer Secretory Vesicles Site Small Interfering RNA Staging Structure Testing Viral Viral Genome Virion Virus Wit Woman cellular targeting cyclophilin B ds-DNA extracellular fighting human PHEMX protein inhibitor/antagonist knock-down mutant neutralizing monoclonal antibodies particle prevent programs public health relevance receptor small molecule tool transcription factor PML uptake

项目摘要

DESCRIPTION (provided by applicant): Human papillomaviruses (HPV) form a large family of viruses some of which are associated with various forms of cancer, including cervical carcinoma. They induce more than seven per cent of all cancers in women worldwide. Especially, HPV pose a burden to women in underdeveloped regions in the world due to a lack of access to cancer screening programs. The long term goal of this research is to understand early events of HPV infection with the focus on events occurring on the cell surface. The major capsid protein, L1, of HPV type 16 is primarily responsible for initial binding of virus to heparan sulfate proteoglycan (HSPG) present on cell surface and extracellular matrix (ECM). Attachment to HSPG induces a shift in the conformation of L1 and the minor capsid protein, L2, both of which we can now detect with monoclonal antibodies. X-ray structure analysis in collaboration with X. Chen identified several HSPG binding sites on the viral capsid. Our preliminary data suggest their sequential use in primary attachment and secondary binding events as well for inducing conformational shifts. In addition, we collected evidence for an involvement of Cyclophilin B (CyPB) in two distinct steps during HPV infection: mediating L2 conformational changes on the cell surface and facilitating uncoating of the viral genome. Aim 1 will test our hypothesis that two HS binding sites on the viral capsid are sequentially used during cell surface events of HPV infection. This will be achieved by a structure-guided mutagenesis of HS binding sites. Aim 2 is intended to delineate the function of Cyclophilins in HPV infection using siRNA knock down and small molecule inhibitors combined with biochemical and cell biological approaches. Aim 3 is dedicated to understand the processes underlying the conformational shifts affecting both capsid proteins in molecular detail. Understanding these events in detail will allow the development of interfering strategies to fight HPV-induced malignant lesions. PUBLIC HEALTH RELEVANCE: HPV induce a variety of cancers, including cervical carcinoma. We will investigate how the virus exploits host cell factors for attachment to host cells and subsequent events leading to successful infection. This should allow identification of cellular targets for drug therapies to prevent HPV induced cancers.

描述（由申请人提供）：人乳头瘤病毒（HPV）形成了大型病毒家族，其中一些与各种形式的癌症有关，包括宫颈癌。他们诱使全世界女性的所有癌症中有7％以上。特别是，由于缺乏获得癌症筛查计划的机会，HPV对世界欠发达地区的妇女造成了负担。这项研究的长期目标是了解HPV感染的早期事件，重点是细胞表面发生的事件。 HPV 16型的主要衣壳蛋白L1主要负责病毒与细胞表面上存在于细胞表面和细胞外基质（ECM）上的硫酸肝素蛋白聚糖（HSPG）的初始结合。对HSPG的附着会诱导L1和次要衣壳蛋白L2的构象的转移，我们现在可以用单克隆抗体检测到。 X射线结构分析与X. Chen合作确定了病毒式衣壳上的几个HSPG结合位点。我们的初步数据表明，它们在主要附件和次要结合事件中的顺序使用以及诱导构象转移。此外，我们收集了在HPV感染期间在两个不同的步骤中涉及环素B（CYPB）的证据：介导细胞表面上的L2构象变化并促进病毒基因组的脱膜。 AIM 1将检验我们的假设，即在HPV感染的细胞表面事件中，在病毒capsid上的两个HS结合位点依次使用。这将通过HS结合位点的结构引导诱变来实现。 AIM 2旨在使用siRNA击倒和小分子抑制剂结合生化和细胞生物学方法来描述周期五肽在HPV感染中的功能。 AIM 3致力于理解以分子细节影响两种衣壳蛋白的构象转移基础的过程。详细了解这些事件将使干扰策略与HPV引起的恶性病变作斗争。公共卫生相关性：HPV诱导各种癌症，包括宫颈癌。我们将研究该病毒如何利用宿主细胞因子与宿主细胞的附着以及随后导致成功感染的事件。这应允许鉴定药物疗法的细胞靶标，以防止HPV诱导的癌症。