Cellular stress response to the oxidizing effects of bleach

细胞对漂白剂氧化作用的应激反应

基本信息

批准号：
8331725
负责人：
Michael Jeffrey Gray
金额：
$ 5.22万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8331725
关键词：
Atherosclerosis Bacteria Biochemical Cells Cellular Stress Response Cellular Structures Chronic Cysteine Disinfectants Disulfides Enzymes Epithelium Escherichia coli Ethylmaleimide Gel Gene Expression Gene Targeting Genetic Techniques Goals Health Household Human Hypochlorous Acid Immune response Immune system In Vitro Inflammation Lactoylglutathione Lyase Leukocytes Malignant Neoplasms Mass Spectrum Analysis Mediating Medical Methods Operon Organism Oxidation-Reduction Oxidative Stress Oxidoreductase Pathogenesis Pathway interactions Peroxides Play Production Proteins Pyruvaldehyde Resistance Reverse Transcriptase Polymerase Chain Reaction Role Sulfhydryl Compounds Superoxides Testing Toxic effect Transcription Repressor/Corepressor Up-Regulation Variant Work acid stress antimicrobial biological adaptation to stress derepression dimer disulfide bond drinking water human disease in vivo insight oxidation oxidative damage protein aggregation repaired research study response

项目摘要

DESCRIPTION (provided by applicant): Hypochlorous acid (HOCl), the active component of household bleach, functions as a powerful antimicrobial during the innate immune response. Despite its widespread use, surprisingly little is known about how bacteria sense and respond to bleach. To our knowledge, no HOCl-sensitive transcriptional regulator has been identified in any organism to date. Preliminary results from our lab implicate the conserved NemR protein as an HOCl-specific transcriptional repressor in E. coli. We have evidence that oxidation by HOCl causes the formation of specific intermolecular disulfide bonds in NemR, which modulate transcriptional activity and induce the expression of genes required for bleach tolerance. I will use both in vitro biochemical approaches and in vivo genetic techniques to examine the role that NemR plays in bacterial bleach resistance. I will investigate the mechanism of HOCl sensing by analyzing the role of conserved cysteine residues in NemR and the structural and functional changes in NemR that occur upon HOCl-treatment. The results of this study will have potentially broad implications on our understanding of how cells sense HOCl and repair cellular damage. Excessive HOCl-production is implicated in the progression of many human diseases, including chronic inflammation, atherosclerosis and cancer and in bacterial interactions with the human immune system.

描述（由申请人提供）：家用漂白剂的活性成分次氯酸酸（HOCL）在先天免疫反应期间起强大的抗菌剂。尽管它广泛使用，但令人惊讶的是，对细菌的感觉和对漂白剂的反应鲜为人知。据我们所知，迄今为止，在任何生物体中尚未确定对HOCL敏感的转录调节剂。我们实验室的初步结果暗示了保守的NEMR蛋白是大肠杆菌中的HOCL特异性转录阻遏物。我们有证据表明，通过HOCL氧化会导致NEMR中特定分子间二硫键的形成，这会调节转录活性并诱导漂白耐受性所需的基因表达。我将使用体外生化方法和体内遗传技术来检查NEMR在细菌漂白耐药性中的作用。我将通过分析保守的半胱氨酸残基在NEMR中的作用以及HOCL治疗时发生的NEMR的结构和功能变化来研究HOCL感测的机制。这项研究的结果将对我们对细胞如何感知HOCL和修复细胞损伤的理解具有广泛的影响。过多的HOCL产生与许多人类疾病的进展有关，包括慢性炎症，动脉粥样硬化和癌症以及与人免疫系统的细菌相互作用。