Subproject 3: Unique Features of Enterococcus that Confer Intrinsic Resistance

子项目 3：肠球菌赋予内在耐药性的独特特征

• 批准号: 10327904
• 负责人: Michael S Gilmore
• 金额: $ 41.12万
• 依托单位: MASSACHUSETTS EYE AND EAR INFIRMARY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10327904
• 关键词: Abnormal Cell; Affect; Animals; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Appearance; Behavior; Biocide; Bioinformatics; Biology; Cell Size; Cell Wall; Cells; Chromosomes; Collection; Communities; Data; Defect; Desiccation; Disinfectants; Distant; Drug Design; Drug Screening; Elements; Enterococcus; Enterococcus faecalis; Enterococcus faecium; Environment; Exposure to; Filament; Gene Silencing; Genes; Genetic; Glean; Goals; Growth; Homeostasis; Hospitals; Image; Image Analysis; Imaging technology; Individual; Infection; Insertion Mutation; Libraries; Microbe; Microfluidic Microchips; Modeling; Modification; Molecular; Morphology; Mothers; Multi-Drug Resistance; Nature; Noise; Nosocomial Infections; Organism; PAWR protein; Pathogenesis; Pathway Analysis; Patients; Phenotype; Play; Population; Process; Property; Proteins; Proteome; Proteomics; Publishing; Recombinants; Regulation; Resistance; Resource Development; Rifampin; Role; Sea; Shapes; Site; Starvation; Stress; Technology; Testing; Time; Variant; antimicrobial; bioinformatics network; cellular imaging; comparative; comparative genomics; defined contribution; fitness; functional genomics; insight; laser tweezer; microfluidic technology; mutant; novel therapeutics; programs; quorum sensing; targeted treatment; technological innovation; trait; transcription factor; transcriptome; transcriptomics; transposon sequencing

SUMMARY The enterococci are ancient microbes that appear to have co-evolved along with the spread of animals on land. They possess an intrinsic ruggedness that allows them to survive unusually harsh conditions, including starvation and desiccation, yet efficiently recolonize the gut of a new host. These traits distinguish them from their ancestors, which led to the identification of an initial set of enterococcal-specific genes and phenotypes. Functional analysis by Tn-seq recently implicated a subset of those genes is important for growth under non-challenged conditions, with still others assuming importance upon antibiotic challenge. This subproject proposes to take advantage of the power of high throughput single cell imaging to more rigorously identify genes critical to the hardiness of enterococci, and associate them with precisely defined growth defects. Moreover it expands this to include genes of importance to various probes of the cell wall, including innate defense molecules, and in the process it will generate an ordered array of known transposon insertion mutations in these genes of importance. As most of the genes that encode the distinctive ruggedness of enterococci encode proteins of unknown function, these mutants will be of considerable value for discovering the underlying mechanisms. Genes found to be important are bioinformatically organized into putative functional networks, which create models that can be tested experimentally. One such network emerged from preliminary studies, and incorporates functions of three critical enterococcal genes with limited distribution outside of the species, that encode genes of unknown function. Bioinformatic analysis suggests that these genes play important roles in regulation of cell wall homeostasis, and appear to be influenced by a quorum sensing mechanism. From functional analysis, this network appears to be highly sensitive to the influence of rifampicin, supporting the role of several key inferred transcription factors in the network. This network will be defined by creating tightly controlled gene silencing constructs and identifying elements of the network through comparative transcriptomics and proteomics. Because of the apparent involvement of three unique enterococcal genes in this network, it appears to be a promising target for limiting the growth and survival of enterococci in patients or in the hospital environment. It is expected that new insights gleaned from the applying Mother Machine imaging technology will add new components to these networks with rigorously defined contributions to enterococcal cell fitness.

概括 肠球菌是一种古老的微生物，似乎是随着细菌的传播而共同进化的。 陆地上的动物。它们具有内在的坚固性，使它们能够异常生存 恶劣的条件，包括饥饿和干燥，但有效地重新殖民新的肠道 主持人。这些特征将他们与祖先区分开来，从而确定了最初的 一组肠球菌特异性基因和表型。最近通过Tn-seq进行功能分析 暗示这些基因的一个子集对于非挑战条件下的生长很重要， 还有一些人认为抗生素挑战很重要。本子项目拟采取 利用高通量单细胞成像的能力来更严格地识别基因 对肠球菌的抵抗力至关重要，并将其与精确定义的生长缺陷联系起来。 此外，它将其扩展为包括对细胞壁的各种探针重要的基因， 包括先天防御分子，在此过程中它将生成已知的有序阵列 转座子插入突变对这些基因具有重要意义。由于大多数编码基因 肠球菌独特的坚固性编码了未知功能的蛋白质，这些突变体 对于发现潜在机制具有重要价值。发现基因 重要的是生物信息学上组织成推定的功能网络，从而创建模型 可以通过实验进行测试。初步研究中出现了一个这样的网络，并且 整合了三种关键肠球菌基因的功能，这些基因在体外分布有限 编码未知功能基因的物种。生物信息分析表明这些 基因在细胞壁稳态的调节中发挥重要作用，并且似乎受到以下因素的影响 群体感应机制。从功能分析来看，这个网络似乎是高度 对利福平的影响敏感，支持几个关键的推断转录的作用 网络因素。该网络将通过创建严格控制的基因沉默来定义 通过比较转录组学构建和识别网络的元素 蛋白质组学。由于三种独特的肠球菌基因明显参与此过程 网络，它似乎是限制肠球菌生长和存活的有希望的目标 患者或医院环境中。预计从应用中收集到新的见解 母机成像技术将严格向这些网络添加新组件 对肠球菌细胞适应性的明确贡献。