Polysomes & Subunits: Structure-Function Relationships

多核糖体

• 批准号: 8241064
• 负责人: ALBERT E DAHLBERG
• 金额: $ 65.73万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1975
• 资助国家: 美国
• 起止时间: 1975-09-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8241064
• 关键词: Address; Amino Acyl Transfer RNA; Antibiotic Resistance; Antibiotics; Anticodon; Applications Grants; Area; Bacteria; Biochemical; Biochemistry; Biological Models; Chemicals; Codon Nucleotides; Complex; Crystallization; Dependence; Development; Drug Design; EEF1A1 gene; Enzymatic Biochemistry; Genetic; Goals; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Hydrolysis; Kinetics; Laboratories; Lead; Life; Medical; Messenger RNA; Methods; Modification; Molecular; Molecular Conformation; Mutation; Pathway interactions; Polyribosomes; Process; Protein Biosynthesis; Proteins; Research; Resistance; Resolution; Ribosomes; Series; Signal Pathway; Signal Transduction; Streptomycin; Structure; Structure-Activity Relationship; Thermus thermophilus; Time; Transfer RNA; X-Ray Crystallography; antimicrobial drug; base; clinically relevant; conformational conversion; design; fighting; genetic analysis; genetic manipulation; insight; interdisciplinary approach; microbial; microorganism; mutant; novel; pathogen; protein complex; public health relevance; research study; resistance mutation; ribosomal protein S12; stereochemistry; structural biology; success; thermophilic bacteria; working group

DESCRIPTION (provided by applicant): ABSTRACT: The long-term goal of this grant proposal is to understand at an atomic level how the ribosome functions during protein synthesis. We will take a multidisciplinary approach utilizing genetic, biochemical and structural methods to study the bacterial ribosome. Our first aim is to examine ribosome structural dynamics during decoding of mRNA and its inhibition by antibiotics. We will use X-ray crystallography to obtain high-resolution structures of mutant Thermus thermophilus 30S subunits that reveal new conformational states. We will examine the effects of the mutations on the accuracy and kinetics of decoding. Our second aim is to probe a pathway by which codon recognition is signaled via ribosomal protein S12 and aminoacyl-tRNA to EF-Tu to activate GTP hydrolysis, a key step in the tRNA selection process. These experiments are designed to provide fundamental insights into ribosome structure and function, including new conformational states of the ribosome, to aid in understanding the decoding process. The ability to solve structures of ribosomes with antibiotic-resistance mutations will aid in understanding the mechanisms of antibiotic action and resistance, and has particular clinical relevance for the rational design of drugs to fight resistant microbial pathogens. PUBLIC HEALTH RELEVANCE: PROJECT NARRATIVE: The goal of this project is to further our understanding of the molecular mechanisms of protein synthesis at the atomic level, using the methods of structural biology, genetics and biochemistry. Protein synthesis in bacteria is the target of over half of all antibiotics, and modifications and mutations in components involved of this process can lead to antibiotic resistance, one of the most pressing medical problems of our time. Deciphering the molecular basis for antibiotic resistance will provide important information needed for the rational design of novel antimicrobial agents effective against antibiotic resistant microorganisms. A more comprehensive examination of the structural basis for protein synthesis will also provide important insights into one of the most fundamental processes found in all forms of life.

描述（由申请人提供）：摘要：这项资助计划的长期目标是在原子水平上了解核糖体在蛋白质合成过程中的功能。我们将采取多学科的方法，利用遗传学，生物化学和结构的方法来研究细菌核糖体。我们的第一个目标是研究核糖体结构的动态解码的mRNA和抗生素的抑制。我们将使用X射线晶体学来获得突变嗜热栖热菌30 S亚基的高分辨率结构，揭示新的构象状态。我们将研究突变对解码的准确性和动力学的影响。我们的第二个目标是探索一条途径，通过该途径，密码子识别通过核糖体蛋白S12和氨酰-tRNA向EF-Tu发出信号，以激活GTP水解，这是tRNA选择过程中的关键步骤。这些实验旨在提供对核糖体结构和功能的基本见解，包括核糖体的新构象状态，以帮助理解解码过程。解决具有耐药性突变的核糖体结构的能力将有助于理解抗生素作用和耐药性的机制，并且对于合理设计药物以对抗耐药性微生物病原体具有特别的临床意义。 公共卫生关系：项目叙述：该项目的目标是利用结构生物学、遗传学和生物化学的方法，进一步了解原子水平上蛋白质合成的分子机制。细菌中的蛋白质合成是超过一半的抗生素的目标，并且该过程所涉及的组分的修饰和突变可能导致抗生素耐药性，这是我们这个时代最紧迫的医学问题之一。解读抗生素耐药性的分子基础将为合理设计有效对抗抗生素耐药性微生物的新型抗菌剂提供重要信息。对蛋白质合成的结构基础进行更全面的研究，也将为了解所有生命形式中最基本的过程之一提供重要的见解。