Biochemistry of lysyl-tRNA synthetases

赖氨酰-tRNA 合成酶的生物化学

• 批准号: 8262939
• 负责人: MICHAEL IBBA
• 金额: $ 31.26万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8262939
• 关键词: Address; Affect; Amino Acids; Amino Acyl Transfer RNA; Amino Acyl-tRNA Synthetases; Aminoacylation; Anthrax disease; Anti-Infective Agents; Antibiotic Resistance; Antibiotic Therapy; Bacteria; Bacterial Model; Binding; Biochemistry; Biological; Biological Models; Catalytic Domain; Cells; Codon Nucleotides; Collaborations; Communicable Diseases; Critical Pathways; Development; Disease; Elongation Factor; Enzymes; Family; Genetic; Goals; Homologous Gene; Lipids; Lysine; Lysine-Specific tRNA; Lysine-tRNA Ligase; Membrane; Membrane Lipids; Messenger RNA; Modification; Molecular; Pathway interactions; Phosphatidyl glycerol; Play; Post-Translational Protein Processing; Process; Property; Protein Biosynthesis; Protein Family; Proteins; Proteome; RNA, Transfer, Amino Acid-Specific; Reaction; Ribosomes; Role; Salmonella; Salmonella infections; Shapes; Specificity; Structure; Testing; Transfer RNA; Translating; Translation Initiation; Translations; Tuberculosis; Virulence; Work; aminoacid biosynthesis; base; factor EF-P; in vivo; insight; member; microbial; paralogous gene; pathogen; trait

DESCRIPTION (provided by applicant): The aminoacyl-tRNA synthetases are responsible for pairing amino acids and tRNAs, and are crucial both for ribosomal protein synthesis and other processes outside translation. The aminoacyl-tRNA synthetase family also contains numerous paralogs that recapitulate either the catalytic core or the various appended domains of aminoacyl-tRNA synthetases. In bacteria the canonical lysyl-tRNA synthetase provides substrates both to translate lysine codons and to modify membrane lipids, while the closely related paralog PoxA instead participates in the post-translational modification of elongation factor P, a pathway critical for the virulence of pathogens such as Salmonella. In both instances, the ability of lysyl-tRNA synthetase-type proteins to promote lysylation of molecules other than tRNA has widespread effects on the cell, determining key properties such as antibiotic resistance. The objective of this proposal is to build on our earlier work on the lysyl-tRNA synthetase family to study non-canonical roles of lysyl-tRNA synthetases and their paralogs. Specifically, we will determine: i) The structural and functional basis of lysyl-tRNA's role in lipd modification. ii) The molecular basis of divergent substrate recognition by lysyl-tRNA synthetase and PoxA. iii) The mechanism of action of modified elongation factor P. This work will reveal the molecular mechanisms that allow members of the lysyl-tRNA synthetase family to participate in aminoacylation reactions outside protein synthesis, and will show how the aminoacylation of proteins and lipids can provide these molecules with new and essential biological activities. Understanding the structural and functional features that allow the lysyl-tRNA synthetase family to play such broad roles in the cell will also provide insights into possible new functions in othe aminoacyl-tRNA synthetase protein families, the vast majority of which contain numerous well-conserved paralogs with no known function. PUBLIC HEALTH RELEVANCE: Anthrax, Salmonellosis and Tuberculosis are infectious diseases with potentially fatal sequelae. Lysyl-tRNA synthetase family proteins contribute to the virulence of the causative agents of these and other diseases by modulating antibiotic resistance and other traits through mechanisms which to date have only been minimally defined. The proposed studies will define the roles of lysyl-tRNA synthetase and its paralog PoxA in establishing antibiotic resistance, and help determine how the corresponding pathways can be used as targets for the development of new anti-infective agents for the treatment of bacterial infectious diseases.

描述（由申请人提供）：氨基酰基-TRNA合成酶负责配对氨基酸和TRNA，对于核糖体蛋白质合成和翻译以外的其他过程至关重要。氨基酰基-TRNA合成酶家族还包含许多旁系同源物，可概括催化核心或氨基酰基-TRNA合成酶的各种附加结构域。在细菌中，规范的赖氨酸-TRNA合成酶既可以提供底物来翻译赖氨酸密码剂和修饰膜脂质，而密切相关的旁系同源物痘则参与了伸长因子P的翻译后修饰，这是病原体诸如鲑鱼类病原体的毒力的关键途径。在这两种情况下，赖氨酸-TRNA合成酶蛋白促进tRNA以外的其他分子的裂解化的能力对细胞具有广泛的影响，从而确定了诸如抗生素耐药性之类的关键特性。该提案的目的是建立我们对赖氨酸-TRNA合成酶家族的早期工作，以研究晶体基-TRNA合酶及其旁同产的非典型作用。具体而言，我们将确定：i）赖氨酸-TRNA在LIPD修饰中作用的结构和功能基础。 ii）赖氨酸-TRNA合成酶和POXA的分子底物识别的分子基础。 iii）修饰伸长因子的作用机理。这项工作将揭示允许赖氨酸-TRNA合成酶家族成员参与蛋白质合成以外的氨基酰胺反应的分子机制，并将显示蛋白质和脂质的氨基糖化如何提供这些分子具有新的分子和新的生物学活性。了解允许赖氨酸-TRNA合成酶家族在细胞中发挥如此广泛的作用的结构和功能特征还将洞悉OTHE氨基酰基-TRNA合成酶蛋白质家族的可能新功能，其中绝大多数包含许多众多保存良好的旁系同源物，却没有已知的功能。 公共卫生相关性：炭疽，沙门氏菌病和结核病是具有潜在致命后遗症的传染病。赖氨酸-TRNA合成酶家族蛋白通过调节抗生素耐药性和其他性状通过迄今为止仅定义的机制来促进这些疾病和其他疾病的致病剂的毒力。拟议的研究将定义赖氨酸-TRNA合成酶及其旁系同源物痘在建立抗生素耐药性中的作用，并有助于确定如何将相应的途径用作开发新抗感染剂以治疗细菌感染疾病的靶标。