New mechanisms of sulfur sensing and trafficking in Staphylococcus aureus.

金黄色葡萄球菌硫传感和运输的新机制。

基本信息

批准号：
8242700
负责人：
DAVID P. GIEDROC
金额：
$ 26.82万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
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：
2011-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8242700
关键词：
2-thiouridine Affinity Alkanesulfonates Anabolism Antibiotic Therapy Area Assimilations Bacteria Binding Biochemical Biogenesis Biological Process Cells Chemicals Community Hospitals Community-Acquired Infections Copper Cysteine Dioxygenases Disease Disulfides Essential Amino Acids Genes Genetic Transcription Homeostasis Hospitals Human Hydrogen Sulfide In Vitro Incidence Infectious Skin Diseases Inorganic Sulfates Intervention Iron-Sulfur Proteins Kinetics Label Low Prevalence Mammals Mass Spectrum Analysis Mediating Mental Depression Metabolic Metabolic Pathway Metabolism Methicillin Resistance Methionine Minor Modeling Modification Molecular Genetics Mycobacterium tuberculosis NAD(P)H dehydrogenase (quinone) 1, human Operon Phenotype Physiologic pulse Proteins Proteomics Regulation Regulon Repression Resistance Resolution Role Sodium Source Staphylococcus aureus Structure Sulfhydryl Compounds Sulfides Sulfites Sulfur Sulfur Metabolism Pathway System Testing Therapeutic Thiamine Thioctic Acid Thiosulfate Sulfurtransferase Toxic effect Transcription Repressor/Corepressor Transfer RNA Unspecified or Sulfate Ion Sulfates Vaccines Work antimicrobial base derepression extracellular in vivo insight mutant novel oxidation paralogous gene pathogen persulfides protein folding public health relevance research study resistant strain sulfurtransferase trafficking

项目摘要

DESCRIPTION (provided by applicant): The Gram-positive opportunistic pathogen Staphylococcus aureus is the causative agent of minor skin infections to far more severe hospital and community-acquired diseases. New antibiotic therapies that target novel metabolic pathways are therefore urgently needed and sulfur metabolism represents one such validated antimicrobial and vaccine strategy. Staphylococcus aureus is characterized by a unique thiol metabolism and is strongly restricted in its ability to obtain inorganic sulfur to make cysteine, an essential amino acid. We have discovered a paralog of a copper-sensing transcriptional repressor CsoR (copper-sensitive operon repressor) in S. aureus strain Newman that we denote CstR, for CsoR-like sulfurtransferase repressor. CstR regulates the expression of a novel operon, cst, which encodes a putative sulfite/sulfonate effluxer (TauE) as well as two multidomain proteins that harbor canonical thiosulfate sulfurtransferase or rhodanese domains (CstA, CstB) whose biological functions are unknown. Our specific objectives are to (1) Elucidate the mechanistic basis of CstR-mediated repression and derepression of cst operon transcription; (2) Elucidate the structure and mechanism of the multidomain thiosulfate sulfurtransferase CstA, which we hypothesize harbors three sulfur relay modules that collectively mediate vectorial transfer of cysteine persulfides from donor to acceptor. NMR structural studies and a novel 34S-32SO32- pulse-32S-32SO32- chase experiment are proposed; (3) Identify cellular proteins that transiently carry 34S-persulfide groups originating with extracellular sodium 34S- 32SO3 thiosulfate using a "bottom-up" proteomics approach; and 4) Begin to elucidate the enzymatic activities of CstR-regulated gene products CstB, a putative rhodanese-containing sulfur dioxygenase and SQR, a putative sulfide:quinone oxidoreductase. This project will provide new insights into the mechanisms of sulfur metabolism and resistance to sulfur metabolite toxicity in an important human pathogen. These studies will significantly extend the emerging paradigm of a cysteine persulfide-based sulfur shuttling system in other bacteria to a new metabolic process as a potential novel antimicrobial intervention strategy. PUBLIC HEALTH RELEVANCE: Staphylococcus aureus is a recognized human pathogen and the causative agent of myriad severe hospital and community acquired infections. In this project, we propose studies to investigate the regulation and function of a novel sulfur metabolic operon in S. aureus that may serve as a new target for antimicrobial therapeutics.

描述（由申请人提供）：革兰氏阳性的机会病原体金黄色葡萄球菌是对更严重的医院和社区获得性疾病的轻微皮肤感染的病因。因此，迫切需要针对新代谢途径的新型抗生素疗法，硫代谢代表一种经过验证的抗菌和疫苗策略。金黄色葡萄球菌的特征是独特的硫醇代谢，并在获得无机硫的能力上受到严格限制，使半胱氨酸是一种必需的氨基酸。我们发现了金黄色葡萄球菌菌株Newman中的铜感应转录阻遏物CSOR（铜敏感的操纵子阻遏物），我们表示CSTR，用于CSOR样硫代硫磺酶抑制剂。 CSTR调节了新型操纵子CST的表达，该操纵子CST编码了推定的亚硫酸盐/磺酸盐外排（TAUE），以及具有含有规范硫代硫酸盐硫酸盐硫酸盐转移酶或犀牛结构域（CSTA，CSTA，CSTB）的两种多域蛋白，其生物功能尚不清楚。我们的具体目标是（1）阐明CST操纵子转录的CSTR介导的抑制和消除的机械基础；（2）阐明了多域硫代硫酸盐硫酸盐转移酶CSTA的结构和机制，我们假设这三个硫磺继电器继电器模块，这些模块共同介导了从供体向受体的硫磺硫化物的矢量转移。提出了NMR结构研究和新型34S-32SO32-PULSE-32S-32SO32-追逐实验。（3）鉴定使用“自下而上”蛋白质组学方法的细胞外钠34S-32SO-32SO3硫代硫酸盐，瞬时携带34s-硫化基团的细胞蛋白； 4）开始阐明CSTR调节的基因产物CSTB的酶促活性，CSTB是一种推定的含荷兰德硫的硫二氧酶和SQR，一种推定的硫化物：喹酮氧化还原酶。该项目将提供有关硫代谢机制和对硫代谢物毒性的抗性的新见解。这些研究将显着扩展其他细菌中基于过硫化半硫化物的硫化硫磺系统的新兴范式，以作为一种新的代谢过程，作为一种潜在的新型抗菌干预策略。公共卫生相关性：金黄色葡萄球菌是公认的人类病原体，也是无数严重医院和社区获得感染的病因。在该项目中，我们建议研究研究新型硫代谢作战孢子中的调节和功能，可以作为抗菌治疗剂的新靶标。