Char of Phospholipases C & the Twin Arginine Secretory System of B. pseudomallei

磷脂酶 C 的炭

• 批准号: 7641024
• 负责人: Michael L. Vasil
• 金额: $ 22.14万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7641024
• 关键词: Affect; Affinity; Anabolism; Archives; Arginine; Bacillus anthracis; Bacteriology; Binding; Biological; Biological Assay; Boxing; Burkholderia mallei; Burkholderia pseudomallei; Candidate Disease Gene; Cell Line; Cell physiology; Cells; Characteristics; Class; Classification; Clinical; Collection; Complement; Conflict (Psychology); DNA Microarray Chip; DNA Microarray format; Deoxyribonuclease I; Deoxyribonucleases; Development; Electrophoretic Mobility Shift Assay; Elements; Endopeptidases; Environment; Equus caballus; Eukaryotic Cell; Evaluation; Evolution; Extracellular Protein; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Genome; Genomics; Glanders; Growth; Hand; Homeostasis; Homologous Gene; Human; In Vitro; Individual; Infection; Intake; Iowa; Iron; Iron-Binding Proteins; Knock-out; Lactoferrin; Learning; Life Style; Ligands; Lipids; Malleus; Mammals; Manuscripts; Metabolism; Methods; Microbe; Microbial Biofilms; Microbial Genetics; Modeling; Molecular Genetics; Mycobacterium tuberculosis; Nitrogen; Nucleic Acids; Nutrient; Organism; Oxidative Stress; Oxygen; Pathogenesis; Peptide Hydrolases; Phenotype; Phospholipase C; Play; Preparation; Process; Prokaryotic Cells; Proteins; Proteomics; Protocols documentation; Pseudomonas aeruginosa; Pseudomonas aeruginosa toxA protein; Purpose; RNase protection assay; Rate; Regulation; Regulator Genes; Regulon; Relative (related person); Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Salmonella typhi; Side; Siderophores; Sigma Factor; Small RNA; Soil; Source; Structural Genes; Surface; System; Therapeutic; Therapeutic Agents; Time; Toxin; Transferrin; Twin Multiple Birth; Universities; Vaccines; Virulence; Virulent; Water; extracellular; heme-binding protein; human tissue; in vivo; interest; macromolecule; macrophage; member; mutant; novel; pathogen; prevent; promoter; prophylactic; siderophore receptors; trafficking; uptake

Introduction The dynamic control of intracellular iron concentrations is paramount to all biological systems. One aspect of this issue is that biologically useful iron (i.e. Fe2") is extremely limiting or it is highly insoluble (i.e.Fe3). Accordingly, biological entities have evolved efficient mechanisms to acquire this nutrient from the insoluble form, which is generally in plentiful quantities. On the other hand, further acquisition of iron above biologically useful concentrations can have dire consequences for a cell. Excess free iron will catalyze the generation of highly reactive oxygen and nitrogen intermediates that will damage all known biological macro-molecules. This conflict, in a major way is dealt with in a diverse array of pathogenic microbes, by a classic Fe2+-dependent represser called Fur (ferric uptake regulator) that plays the key role in controlling iron homeostasis. While it is clear that the Class B Select Agents Burkholderia pseudomallei & mallei express a protein that is highly homologous to Fur from closely related organisms (e.g. Pseudomonas aeruginosa) very little is known about the role of iron homeostasis in the virulence of 8. pseudomallei & mallei. More importantly, nothing is known about the role of iron homeostasis in the intracellular life style of these pathogens. We propose to remedy this situation through the use of powerful molecular and genetic approaches that we employed to elucidate the extensive and crucial role that Fur plays in the related, but distinct, opportunistic pathogen P. aeruginosa. We will identify the Fur regulated genes of 6. pseudomallei & mallei that encode regulatory factors (e.g. sigma factors, small regulatory RNAs). We will then evaluate the role of these selected regulatory factors in the ability of these agents to enter, survive in, grow in or traffick in model eukaryotic cells (e.g. macrophage), which are appropriate to the known aspects of the pathogenesis of these potential bioterrorist agents. The gene products that we identify may ultimately be used in developing therapeutic and prophylactic agents against Malleipdosis and Glanders. Project interactions In addition to the obvious interactions with other project members (e.g. Drs. Vazquez- Torres, Holmes, & Voskuil) that are described in more detail in the body of this application and in the Introduction to Project II.C, we will especially interact with Dr. Robison (BYU Select Agent Archive) and Dr. Schweizer (Microbial genetics - CSU) in terms of the bacteriology and genetics of B. pseudomallei & mallei. We will also be especially interested in very active interactions with Dr. Slayden (Genomics/Proteomics Core) that will be vital to development of microarrays and for the evaluation of potential target genes and products for possible use in experimental prophylactic and therapeutic agents. Also, along these lines we will also closely interact with Dr. Brennan (PDM Core) for the same purpose.

简介细胞内铁浓度的动态控制对于所有生物系统至关重要。 这个问题的一个方面是，生物学上有用的铁（即Fe2”）极为局限，或者它是高度不溶性的 （即fe3）。因此，生物实体已经发展了有效的机制，从 不溶性形式，通常数量很多。另一方面，在上面进一步获取铁 生物学上有用的浓度可能会对细胞产生可怕的后果。多余的自由铁会催化 产生高反应性氧和氮中间体，这些中间体会损害所有已知的生物学 宏观分子。这种冲突以一种主要的方式在各种各样的致病微生物中处理 经典Fe2+依赖性阻遏物称为皮毛（铁摄取调节剂），在控制铁中起关键作用 稳态。虽然很明显，B级选择代理商Burkholderia pseudomallei＆Mallei表示 与紧密相关的生物（例如铜绿假单胞菌）高度同源的蛋白质非常同源 关于铁体内平衡在8。假单胞菌和马利的毒力中的作用知之甚少。更多的 重要的是，关于铁稳态在细胞内生活方式中的作用一无所知 病原体。我们建议通过使用强大的分子和遗传来解决这种情况 我们采用的方法来阐明毛皮在相关中所扮演的广泛和关键作用，但 独特的机会性病原体铜绿假单胞菌。我们将确定6个毛皮调节基因。 编码调节因子的MALLEI（例如Sigma因子，小监管RNA）。然后我们将评估 这些选定的调节因素在这些药物进入，生存，生存或贩运的能力中的作用 模型真核细胞（例如巨噬细胞），适合发病机理的已知方面 在这些潜在的生物恐怖剂中。我们识别的基因产品最终可以用于 开发针对疟原虫和腺体的治疗和预防剂。 除了与其他项目成员的明显互动之外，项目相互作用（例如Vazquez-博士 托雷斯，福尔摩斯和沃斯库尔）在本应用的正文中更详细地描述 II.C项目简介，我们将特别与Robison博士（BYU Select Agent Archive）和博士进行互动。 Schweizer（微生物遗传学-CSU）就假杆菌和马利芽孢杆菌的细菌学和遗传学而言。 我们还将对与Slayden博士非常活跃的互动特别感兴趣（基因组学/蛋白质组学 核心）将对微阵列的发展以及对潜在靶基因和潜在靶基因的评估至关重要 可能用于实验性预性和治疗剂的产品。另外，沿着这些线路我们将 出于相同的目的，也与Brennan博士（PDM Core）紧密互动。