Biosynthesis & Function of a Bacillus anthracis-specific cell wall polysaccharide

生物合成

基本信息

批准号：
7739417
负责人：
RUSSELL W CARLSON
金额：
$ 22.26万
依托单位：
UNIVERSITY OF GEORGIA
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：
2009-06-05 至 2011-05-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7739417
关键词：
Address Affect Anabolism Animals Anthrax disease Antibodies Antigens Autolysin Bacillus (bacterium)Bacillus anthracis Bacillus anthracis spore Bacillus cereus Bacteria Bacteriophages Binding Binding Proteins Bioterrorism Carbohydrates Cell Surface Proteins Cell Wall Cell surface Cells Data Deletion Mutation Development Diagnostic Disease Engineering Enzymes Evaluation Exposure to Funding Future Gene Expression Genes Genome Growth Growth and Development function Human Hydrogen Fluoride Immune Sera Institutes Investigation Knowledge Laboratories Letters Life Literature Macaca mulatta Measures Medical Research Mutagenesis Mutation Parents Pathogenicity Pathway interactions Peptidoglycan Phenotype Physiological Plasmids Polymers Polysaccharides Preparation Primates Property Prophages Proteins Public Health Public Health Practice Publications Regulation Reporting Research Role Serum Staging Structure Temperature Therapeutic Transferase Transferase Gene UDP-N-acetylglucosamine 2-epimerase Vaccinated Vaccines Virulence Virulence Factors Work anti-IgG aqueous base cell envelope cell growth endolysin falls gene synthesis immunogenic improved insight lysin mouse model mutant novel public health relevance vaccine development

项目摘要

DESCRIPTION (provided by applicant): Our laboratory has recently reported the structure of a Bacillus anthracis species-specific polysaccharide. This polysaccharide (termed HF-PS) has been classified a 'non-classical' secondary cell wall polymer. It is isolated from the cell wall of vegetative cells by treatment with aqueous hydrogen fluoride (HF), and was previously reported to function in anchoring/exporting the S-layer proteins, Sap and EA1, to the cell surface. Our work strongly supports that the HF-PS is structurally specific to B. anthracis strains, is immunogenic in that animals injected with live or dead spores of B. anthracis Sterne (34F2) produce antibodies against HF-PS, and is immunogenically specific with regard to the binding of these antibodies to the HF-PSs of even closely related B. cereus strains. Further, we have shown that sera from Rhesus macaques that survive exposure to B. anthracis spores contain IgG anti-HF-PS antibodies. Preliminary data and results in the literature support that these polysaccharides could be crucial for important functions with regard to the growth and virulence of pathogenic Bacillus species, including B. anthracis. However, to date direct empirical evidence is still missing. The aims of this proposal are to engineer mutants that are impaired in the biosynthesis of this HF-PS polysaccharide and to characterize the phenotype of these mutants with regard to their growth, sporulation, HF-PS structure, HF-PS interaction with S-layer proteins and various phage endolysins, and to study the effect of each mutation on the virulence in a mouse model. This research will provide us with necessary insight regarding the HF-PS's pathogenic functions, and its usefulness for the development of vaccines, diagnostics and therapeutics. PUBLIC HEALTH RELEVANCE: The bacterium Bacillus anthracis causes anthrax in humans; as a bioterrorism agent the bacteria are a threat to public health. Our laboratory has recently reported a cell wall polysaccharide of a B. anthracis (termed HF- PS) with a structure specific to these bacteria. We will create B. anthracis mutants impaired in the biosynthesis of this cell wall polysaccharide and investigate whether these mutants are impaired in important bacterial growth functions and disease development. This investigation will lay the groundwork for future studies into improved vaccines, diagnostics, and therapeutics, and into new anthrax-related public health practices.

描述（由申请人提供）：我们的实验室最近报道了炭疽芽孢杆菌物种特异性多糖的结构。这种多糖（称为 HF-PS）已被归类为“非经典”次生细胞壁聚合物。它是通过氟化氢 (HF) 水溶液处理从营养细胞的细胞壁中分离出来的，之前有报道称它具有将 S 层蛋白 Sap 和 EA1 锚定/输出到细胞表面的功能。我们的工作强烈支持 HF-PS 在结构上对炭疽芽孢杆菌菌株具有特异性，具有免疫原性，因为注射炭疽芽孢杆菌 Sterne (34F2) 活孢子或死孢子的动物会产生针对 HF-PS 的抗体，并且在以下方面具有免疫原特异性：这些抗体与甚至密切相关的蜡状芽孢杆菌菌株的 HF-PS 的结合。此外，我们还发现，暴露于炭疽芽孢杆菌孢子后仍存活的恒河猴血清中含有 IgG 抗 HF-PS 抗体。文献中的初步数据和结果表明，这些多糖对于致病性芽孢杆菌（包括炭疽芽孢杆菌）的生长和毒力的重要功能至关重要。然而，迄今为止，仍然缺乏直接的经验证据。该提案的目的是设计在这种 HF-PS 多糖生物合成中受损的突变体，并表征这些突变体的表型，包括其生长、孢子形成、HF-PS 结构、HF-PS 与 S 层的相互作用蛋白质和各种噬菌体内溶素，并研究每种突变对小鼠模型毒力的影响。这项研究将为我们提供有关 HF-PS 致病功能及其对开发疫苗、诊断和治疗的有用性的必要见解。公共卫生相关性：炭疽杆菌会引起人类炭疽病；作为一种生物恐怖主义媒介，细菌对公众健康构成威胁。我们的实验室最近报道了炭疽芽孢杆菌的细胞壁多糖（称为 HF-PS），其具有这些细菌特有的结构。我们将创建这种细胞壁多糖生物合成受损的炭疽芽孢杆菌突变体，并研究这些突变体的重要细菌生长功能和疾病发展是否受损。这项调查将为未来改进疫苗、诊断和治疗方法以及与炭疽相关的新公共卫生实践的研究奠定基础。