Cell separation in Neisseria gonorrhoeae

淋病奈瑟菌的细胞分离

基本信息

批准号：
8417663
负责人：
Joseph P Dillard
金额：
$ 22.58万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-05 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8417663
关键词：
Affect Amidohydrolases Animal Model Antibiotic Resistance Antibiotic Therapy Antibiotics Antibodies Autolysis Bacillus subtilis Bacteria Biochemical Biochemical Reaction Biological Assay Cell Fraction Cell Separation Cell Wall Cells Cervical Characteristics Combined Modality Therapy Complex DNA Data Defect Detergents Disease Ectopic Pregnancy Electron Microscopy Enzyme Inhibitor Drugs Enzyme Inhibitors Enzyme Interaction Enzymes Escherichia coli Future Gene Deletion Genes Gonorrhea Growth Human Immunoprecipitation In Vitro Infection Infertility Inflammatory Response Invaded Knowledge Location Lytic Measures Membrane Metabolism Methods Microtomy Multi-Drug Resistance Mutation Neisseria gonorrhoeae Normal Cell Pelvic Inflammatory Disease Peptidoglycan Population Process Proteins Role Scaffolding Protein Scheme Sexually Transmitted Diseases Shapes System Testing Time Western Blotting Woman amidase antimicrobial base combat drug sensitivity in vitro Assay inhibitor/antagonist link protein men mutant novel pathogen prevent research study resistant strain

项目摘要

DESCRIPTION (provided by applicant): Neisseria gonorrhoeae causes a very common sexually transmitted disease in men and women that often results in severe consequences for women including pelvic inflammatory disease, ectopic pregnancy, and infertility. The gonococcal population is becoming increasingly antibiotic resistant, and multidrug resistant strains have arisen. This proposal seeks to understand the mechanisms of peptidoglycan breakdown involved in cell separation and to develop cell separation as a target for antimicrobial therapy. Preliminary data indicate that defects in cell separation make N. gonorrhoeae more sensitive to antibiotics as well as defective in infection of human cells. Previously we demonstrated that mutations in genes for peptidoglycan degradation - amiC or ltgC - in N. gonorrhoeae cause aberrant growth, loss of cell separation, and increased autolysis. These mutants grow in large aggregates sharing a single interconnected cell wall and with a single outer membrane surrounding the aggregate. Here we will 1) determine the biochemical functions of two peptidoglycan-degrading enzymes, AmiC and LtgC, required for cell separation and develop assays that could be used to identify inhibitors of these enzymes, 2) examine the localization and possible interactions of AmiC and LtgC in gonococcal cells in order to determine the function of these enzymes for cell separation and develop assays for these enzymes in cells, and 3) determine the extent of defects of cell separation mutants for infection of human cells and for resistance to antibiotics. Knowledge gained in this study and assays developed here will facilitate the identification of antibiotics for the treatment of gonococcal diseases and for combating other bacteria that have similar mechanisms of cell separation.

描述（由申请人提供）：淋病奈瑟菌在男性和女性中引起一种非常常见的性传播疾病，通常会给女性带来严重的后果，包括盆腔炎、宫外孕和不孕症。淋球菌群体对抗生素的耐药性日益增强，并且出现了多重耐药菌株。该提案旨在了解细胞分离中涉及的肽聚糖分解机制，并将细胞分离开发为抗菌治疗的目标。初步数据表明，细胞分离缺陷使淋病奈瑟菌对抗生素更加敏感，并且在感染人体细胞方面存在缺陷。之前我们证明了淋病奈瑟菌中肽聚糖降解基因（amiC 或 ltgC）的突变会导致异常生长、细胞分离丧失和自溶增加。这些突变体以大聚集体的形式生长，共享单个互连的细胞壁并具有围绕聚集体的单个外膜。在这里，我们将 1) 确定细胞分离所需的两种肽聚糖降解酶 AmiC 和 LtgC 的生化功能，并开发可用于鉴定这些酶抑制剂的检测方法，2) 检查 AmiC 和 LtgC 的定位和可能的相互作用在淋球菌细胞中，以确定这些酶在细胞分离中的功能并开发细胞中这些酶的测定方法，以及 3) 确定细胞分离缺陷的程度用于感染人类细胞和对抗生素产生耐药性的突变体。在这项研究中获得的知识和这里开发的测定将有助于鉴定用于治疗淋球菌疾病和对抗具有类似细胞分离机制的其他细菌的抗生素。