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：
7740732
负责人：
Martin Sapp
金额：
$ 36.63万
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/7740732
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）形成一个大的病毒家族，其中一些与各种形式的癌症相关，包括宫颈癌。全球女性癌症中超过 7% 是由它们诱发的。尤其是，由于无法获得癌症筛查项目，HPV 给世界欠发达地区的女性带来了负担。这项研究的长期目标是了解 HPV 感染的早期事件，重点关注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和细胞外基质 (ECM) 上存在的硫酸乙酰肝素蛋白聚糖 (HSPG) 的初始结合。与 HSPG 的附着会诱导 L1 和次要衣壳蛋白 L2 的构象发生变化，我们现在可以用单克隆抗体检测这两者。与 X. Chen 合作进行的 X 射线结构分析确定了病毒衣壳上的几个 HSPG 结合位点。我们的初步数据表明它们在初级附着和次级结合事件中的顺序使用以及诱导构象转变。此外，我们收集了亲环蛋白 B (CyPB) 参与 HPV 感染过程中两个不同步骤的证据：介导细胞表面 L2 构象变化和促进病毒基因组脱壳。目标 1 将检验我们的假设，即在 HPV 感染的细胞表面事件期间，病毒衣壳上的两个 HS 结合位点顺序使用。这将通过 HS 结合位点的结构引导诱变来实现。目标 2 旨在使用 siRNA 敲低和小分子抑制剂结合生化和细胞生物学方法来描述亲环蛋白在 HPV 感染中的功能。目标 3 致力于了解影响两种衣壳蛋白分子细节的构象变化背后的过程。详细了解这些事件将有助于制定干扰策略来对抗 HPV 引起的恶性病变。公共卫生相关性：HPV 会诱发多种癌症，包括宫颈癌。我们将研究病毒如何利用宿主细胞因子附着于宿主细胞以及导致成功感染的后续事件。这应该能够识别药物治疗的细胞靶标，以预防 HPV 诱发的癌症。