Biochemistry of lysyl-tRNA synthetases

描述(申请人提供)：氨基酰-tRNA合成酶负责配对氨基酸和tRNA，对核糖体蛋白质合成和翻译外的其他过程都是至关重要的。氨基酰基-tRNA合成酶家族还包含大量的副产物，它们概括了氨基酰基-tRNA合成酶的催化核心或各种附加结构域。在细菌中，规范的赖氨酸-tRNA合成酶提供了翻译赖氨酸密码子和修饰膜脂的底物，而密切相关的Paralog PoxA则参与了延伸因子P的翻译后修饰，这是沙门氏菌等病原体毒力的关键途径。在这两种情况下，赖氨酰-tRNA合成酶类型的蛋白质促进tRNA以外的分子裂解的能力对细胞有广泛的影响，决定了抗生素耐药性等关键特性。这项建议的目的是在我们早期关于赖氨酰-tRNA合成酶家族的工作的基础上，研究赖氨酰-tRNA合成酶及其类似物的非典范作用。具体地说，我们将确定：i)赖氨酰-tRNA在脂肪修饰中作用的结构和功能基础。Ii)赖氨酰-tRNA合成酶和PoxA识别发散底物的分子基础。这项工作将揭示赖氨酰-tRNA合成酶家族成员参与蛋白质合成之外的氨基酰化反应的分子机制，并揭示蛋白质和脂类的氨酰化如何为这些分子提供新的和必要的生物活性。了解赖氨酰-tRNA合成酶家族在细胞中发挥如此广泛作用的结构和功能特征，也将有助于深入了解其他氨基酰基-tRNA合成酶蛋白家族可能的新功能，其中绝大多数包含许多保守的、未知功能的旁系蛋白。 与公共卫生相关：炭疽病、沙门氏菌病和结核病是具有潜在致命后遗症的传染病。赖氨酰-tRNA合成酶家族蛋白通过调节抗生素耐药性和其他特征来促进这些和其他疾病的病原体的毒力，到目前为止，这些机制还只被最低限度地定义。这项拟议的研究将确定赖氨酰-tRNA合成酶及其类似的POXA在建立抗生素耐药性方面的作用，并有助于确定如何将相应的途径用作开发治疗细菌感染性疾病的新的抗感染药物的靶点。

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.