Determinants of Pathogenic Hantavirus Attachment

致病性汉坦病毒附着的决定因素

• 批准号: 7860323
• 负责人: Erich R Mackow
• 金额: $ 19.61万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7860323
• 关键词: Antibodies; Antibody Formation; Antiviral Agents; Binding; Binding Sites; Biochemical; Blocking Antibodies; Cleaved cell; Development; Disease; Endothelial Cells; Escape Mutant; Figs - dietary; Glycoproteins; Golgi Apparatus; Hantavirus; Hantavirus Infections; Hantavirus Pulmonary Syndrome; Hemorrhagic Fever with Renal Syndrome; Integrins; Mediating; Membrane Glycoproteins; Membrane Proteins; Peptides; Polyproteins; Protein Binding; Protein Structure Initiative; Proteins; Proteomics; Research; Retroviridae; Role; Surface; Tertiary Protein Structure; Testing; Therapeutic; Vascular Permeabilities; Viral; Viral Proteins; Virus; base; cell motility; crosslink; inhibitor/antagonist; neutralizing antibody; positional cloning; prevent; public health relevance; receptor; receptor binding; research study; therapeutic vaccine; trafficking

DESCRIPTION (provided by applicant): Hantaviruses infect endothelial cells (ECs) and cause 2 vascular permeability-based diseases: Hemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). Pathogenic hantaviruses bind to a 53 residue PSI domain present at the apex of bent, inactive, 1v23 integrins and dysregulate 1v23 integrin function (33, 73). The absence of 1v23 function is a known cause of vascular permeability and hemorrhagic disease, and only pathogenic hantaviruses bind 1v23. Hantavirus binding to 23 is RGD independent, however hantavirus attachment proteins and domains required for binding have not been discovered. We have identified 2 differences between surface proteins of pathogenic and non-pathogenic hantaviruses which correlate with 1v23 integrin usage. Here we will investigate the domains and residues of hantavirus surface glycoproteins required for pathogenic hantavirus binding to the 23 integrin PSI domain. Hantavirus surface proteins are synthesized as a polyprotein that is co-translationally cleaved into Gn and Gc fragments and trafficked to the cis-Golgi where hantavirus budding occurs. To date all pathogenic hantaviruses tested use 1v23 integrins for viral entry while non-pathogenic hantaviruses use discrete 1521 integrins. These findings suggest that unique changes in hantavirus Gn or Gc surface proteins are likely to differentiate viral attachment of pathogenic and non-pathogenic hantaviruses. We identified 2 Gc residue changes that are unique to non-pathogenic hantaviruses and not present in highly divergent pathogenic hantaviruses. Our studies of pathogenic hantavirus binding to 23 integrin PSI domains provide a basis for defining hantavirus proteins, domains and residues that mediate attachment. Proposed studies are likely to identify virus specific targets for the development of hantavirus therapeutics and vaccines and antibodies to viral attachment domains are likely to have direct therapeutic application. Objective: In this proposal we will define the domains and residues of pathogenic hantaviruses required for binding to 23 intergrin PSI domains and determine whether these domains elicit neutralizing antibody responses. Specific Aims: 1) Analyze PSI Domain Binding to Pathogenic Hantavirus Surface Proteins 2) GnGc Pseudotyped Virus Will be used to Define Requirements for 1v23 Binding 3) Antibodies to GnGc Peptides Required for PSI Binding will be evaluated for their ability to Inhibit Hantavirus Infection and PSI Domain Binding PUBLIC HEALTH RELEVANCE: Pathogenic hantaviruses bind to a small protein domain on 23 receptors called the PSI domain and targeting the viral protein that is responsible for binding to the 23 receptor is a viable means of inhibiting hantavirus infectivity and may be an antiviral therapeutic approach for regulating pathogenic hantavirus disease. Here we propose to define the viral protein components that are responsible for attaching the virus to the 23 PSI domain. Small pieces of hantavirus surface proteins will be analyzed for their ability to bind 1v23 PSI domains, block hantavirus infection, elicit antibodies that bind hantaviruses and prevent hantavirus interactions with cellular 1v23 receptors.

描述（由申请方提供）：汉坦病毒感染内皮细胞（EC）并引起2种基于血管通透性的疾病：肾综合征出血热（HFRS）和汉坦病毒肺综合征（HPS）。致病性汉他病毒与存在于弯曲的、无活性的1v23整联蛋白的顶点处的53个残基PSI结构域结合，并使1v23整联蛋白功能失调（33，73）。1v23功能的缺失是血管通透性和出血性疾病的已知原因，并且只有致病性汉坦病毒结合1v23。汉坦病毒与23的结合是不依赖于RGD的，然而尚未发现结合所需的汉坦病毒附着蛋白和结构域。我们已经确定了致病性和非致病性汉坦病毒的表面蛋白之间的2个差异，其与1v23整合素的使用相关。在这里，我们将研究致病性汉他病毒结合23整合素PSI结构域所需的汉他病毒表面糖蛋白的结构域和残基。汉坦病毒表面蛋白合成为多聚蛋白，其共裂解成Gn和Gc片段并运输到汉坦病毒出芽发生的顺式高尔基体。迄今为止，所有检测的致病性汉他病毒都使用1v23整联蛋白进行病毒进入，而非致病性汉他病毒使用离散的1521整联蛋白。这些发现表明汉坦病毒Gn或Gc表面蛋白的独特变化可能区分致病性和非致病性汉坦病毒的病毒附着。我们确定了2个Gc残基的变化，这是独特的非致病性汉他病毒，而不存在于高度分歧的致病性汉他病毒。我们的致病性汉他病毒结合23整合素PSI结构域的研究提供了定义汉他病毒的蛋白质，结构域和残基介导的附件的基础。拟议的研究可能会确定病毒特异性靶点，用于开发汉他病毒治疗剂和疫苗，病毒附着结构域的抗体可能具有直接的治疗应用。目的：在这个提议中，我们将定义致病性汉坦病毒的结构域和残基所需的结合23整合素PSI结构域，并确定这些结构域是否引起中和抗体反应。具体目标：1）分析PSI结构域与致病性汉坦病毒表面蛋白的结合2）GnGc假型病毒将用于定义1v23结合的要求3）评价PSI结合所需的GnGc肽的抗体抑制汉坦病毒感染和PSI结构域结合的能力公共卫生相关性：致病性汉坦病毒与23受体上的一个小蛋白质结构域结合，称为PSI结构域，靶向负责与23受体结合的病毒蛋白质是抑制汉坦病毒的可行方法可能是调节致病性汉坦病毒病的抗病毒治疗方法。在这里，我们建议定义的病毒蛋白质的组成部分，负责连接病毒的23 PSI结构域。将分析小块汉他病毒表面蛋白结合1v23 PSI结构域、阻断汉他病毒感染、引发结合汉他病毒的抗体和阻止汉他病毒与细胞1v23受体相互作用的能力。