Herpes simplex virus receptors and signal transduction

描述(申请人提供)：单纯疱疹病毒(HSV)通过与细胞表面硫酸乙酰肝素链结合而附着在细胞上，然后与其他几种细胞表面分子中的任何一种结合，触发病毒粒子被膜和细胞膜之间的融合，从而导致病毒进入。其他细胞表面受体包括肿瘤坏死因子受体家族成员hvem；免疫球蛋白超家族的细胞黏附分子Nectin-1和Nectin-2；以及由某些3-O-磺基转移酶产生的硫酸乙酰肝素中的特定位点。病毒粒子gD是所有这些进入受体的配基。HSV-1 gD可与HSEM、Nectin-1和3-O-硫酸乙酰肝素结合，而HSV-2 gD可与HVEM、Nectin-1和Nectin-2结合。HSV-1gD(RID)的某些突变形式已经失去了与HVEM和3-O-硫酸乙酰肝素结合的能力，而获得了与Nectin-2结合的能力。父母资助的目的涉及HSV进入人类细胞的要求，重点是进入受体的结构/功能研究，以及它们在HSV进入特定人类细胞类型中的作用。在这个探索性的方案中，我们打算确定HSV-1或HSV-1/Ridl gD是否可以通过任何蛋白质进入受体传递或中断信号，以及不同进入受体的参与是否传递不同的信号。将使用表达HVEM、Nectin-1和Nectin-2的野生型人成纤维细胞和表达Nectin-1的突变成纤维细胞。这些细胞将暴露在外源性HSV-1或HSV-1/RIDL GD中，或将被诱导表达每种形式的GD。将从处理或模拟处理的细胞中分离RNA，通过将标记的CRNAs与DNA寡核苷酸微阵列杂交来比较细胞基因转录水平。我们的假设是，GD介导的信号转导或中断将导致特定细胞mRNAs水平的变化，观察到的具体变化将取决于GD参与的特定受体。如果事实证明是这样，一个长期目标将是确定病毒进入过程中的信号转导是否以受体依赖的方式影响病毒进入后的事件。本提案中描述的研究不同于由母项目资助的研究，但涉及的问题源于母项目中的调查结果。

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) attaches to cells by binding to cell surface heparan sulfate chains and then engages any one of several other cell surface molecules to trigger fusion between the virion envelope and a cell membrane, which leads to viral entry. The other cell surface receptors (entry receptors) include HVEM, a member of the TNF receptor family; nectin-1 and nectin-2, cell adhesion molecules of the Ig superfamily; and specific sites in heparan sulfate generated by certain 3-O-sulfotransferases. Virion gD is the ligand for all of these entry receptors. HSV-1 gD can bind to HVEM, nectin-1 and 3-O-sulfated heparan sulfate whereas HSV-2 gD can bind to HVEM, nectin-1 and nectin-2. Certain mutant forms of HSV-1 gD (Rid) have lost the ability to bind to HVEM and 3-O-sulfated heparan sulfate and acquired the ability to bind to nectin-2. The aims of the parent grant relate to the requirements for HSV entry into human cells and focus on structure/function studies of the entry receptors and characterization of their roles in HSV entry into specific human cell types. In this exploratory proposal, we intend to determine whether HSV-1 or HSV-1/Ridl gD can transduce or interrupt signals via any of the protein entry receptors and whether engagement of different entry receptors transduces different signals. Wild-type human fibroblasts expressing HVEM, nectin-1 and nectin-2 and mutant fibroblasts defective for nectin-1 expression will be used. These cells will be exposed to exogenous HSV-1 or HSV-1/Ridl gD or will be made to express each form of gD. RNAs will be isolated from the treated or mock-treated cells for comparisons of cell gene transcript levels by hybridization of labeled cRNAs to DNA oligonucleotide microarrays. Our hypothesis is that transduction or interruption of signals mediated by gD will result in changes in the levels of specific cell mRNAs and that the specific changes observed will depend on the particular receptor engaged by gD. If this proves to be the case, a long-term goal will be to determine whether signal transduction during viral entry influences post-entry events in viral infection in a receptor-dependent fashion. The studies described in this proposal are distinct from those supported by the parent grant but address questions that arise from findings made in the parent project.