INHIBITORS OF BACTERIAL RNA POLYMERASE: "SWITCH REGION"

细菌 RNA 聚合酶抑制剂：“转换区域”

• 批准号: 7742240
• 负责人: RICHARD H. EBRIGHT
• 金额: $ 58.22万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7742240
• 关键词: Address; Affect; Anti-Bacterial Agents; Bacterial Infections; Bacterial RNA; Binding; Biological Factors; Biological Warfare; Bioterrorism; Clinical; Cloning; Complex; Crystallography; DNA; DNA-Directed RNA Polymerase; Development; Elements; Escherichia coli; Evaluation; Exhibits; Future; Gene Cluster; Genetic Transcription; Growth; Lactones; Mediating; Molecular Cloning; Molecular Conformation; Polymerase; Public Health; Pyrones; RNA Polymerase I; RNA Polymerase II; RNA Polymerase III; Reaction; Research Personnel; Resistance; Rifabutin; Rifampin; Screening procedure; Structure; Thermus thermophilus; Transcription Initiation; Work; base; inhibitor/antagonist; novel; programs; resistant strain; rifapentine; single molecule; single-molecule FRET; small molecule; tool

Preliminary work establishes that three natural products-the polyketide-derived a-pyrone myxopyronin, the Dolyketide-derived a-pyrone corallopyronin, and the polyketide-derived macrocylic lactone ripostatin-inhibit Dacterial RNA polymerase (RNAP) through interactions with the RNAP "switch region," a structural element that mediates conformational changes required for RNAP to bind and retain the DMAtemplate in transcription. The compounds do not inhibit eukaryotic RNAP I, RNAP II, or RNAP III. The compounds ootently inhibit Gram-positive and Gram-negative bacterial growth, exhibit no cross-resistance with the nhibitors of bacterial RNAP in current clinical use in therapy of bacterial infection (therifamycin antibacterial agents, rifampicin, rifapentine, and rifabutin), and exhibit no cross-resistance with other inhibitors of bacterial RNAP under evaluation for future clinical use in therapy of bacterial infection. The proposed work wilt use x-ray crystallography, ensemble and single-molecule fluorescence resonance energy transfer, single-molecule nanomanipulation, molecular cloning, surrogate-host expression, structure-based screening, and de novo screening, to address four specific aims: Specific Aim 1: Determination of structures of complexes of RNAP with switch-region-target inhibitors Specific Aim 2: Determination of mechanisms of inhibition of RNAP by switch-region-target inhibitors Specific Aim 3: Cloning, characterization, and surrogate-host expression of biosynthetic gene clusters for switch-region-target inhibitors Specific Aim 4: Identification and characterization of novel switch-region-target inhibitors The results will enable development of new broad-spectrum antibacterial agents that will be effective against bacterial strains resistant to currently used antibacterial agents. As such, the results will have direct relevance to public health and to development of countermeasures against bacterial strains that could be used in biowarfare or bioterrorism. In addition, the results will contribute to understanding RNAP structure and function and will provide tools for analysis of RNAP structure and function.

初步的工作确定了三种天然产物——聚酮衍生的a-吡咯酮黏性吡咯肽