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Studies on Transfer RNA

转移RNA的研究

基本信息

批准号：
7849876
负责人：
DIETER SOLL
金额：
$ 66.86万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7849876
关键词：
ATP phosphohydrolase Address Affect African Trypanosomiasis Amides Amino Acid Sequence Amino Acids Amino Acyl Transfer RNA Amino Acyl-tRNA Synthetases Aminoacylation Ammonia Anabolism Anti-Infective Agents Antigen Targeting Archaea Area Arginine Asparagine Asparagine-Specific tRNA Autoimmune Hepatitis Behavior Binding Binding Sites Biochemical Biochemical Genetics Biochemistry Biological Assay Biology Cell physiology Cells Charge Chemicals Code Complex Cysteine Cysteine-Specific tRNA DNA DNA Sequence Dependence Discrimination EEF1A1 gene Elongation Factor Engineering Enzymes Escherichia coli Eukaryota Galactosidase Genes Genetic Genetic Code Glutaminase Glutamine Glutamine-Specific tRNA Goals Helicobacter pylori Holoenzymes Housekeeping Human In Vitro Investigation Knock-out Knowledge LacZ Genes Lead Ligase Light Link Lysine Macromolecular Complexes Malaria Malignant Neoplasms Metabolic Metabolic Diseases Methanobacteria Methanococcus Methanothermobacter Methodology Methods Methyltransferase Micronutrients Modification Molecular Multienzyme Complexes Mutagenesis N-terminal Nanoarchaeum Nature Nerve Degeneration Organism Pathway interactions Patients Phosphotransferases Plasmodium falciparum Positioning Attribute Post-Translational Protein Processing Process Protein Biosynthesis Proteins Pyrimidine Nucleotides Quality Control RNA Interference Reaction Ribosomes Role Route Screening procedure Selenium Selenocysteine Serine-tRNA Ligase Shipping Ships Side Structure Sulfur Surveys System Technology Testing Transfer RNA Translation Process Translations Transplantation Trypanosoma brucei brucei Upper arm Urea Variant Work base combat cysteine-tRNA cysteinylcysteine design directed evolution frontier glutamine-tRNA human disease in vivo insight interest mutant novel nucleotide metabolism nutrition pathogen pressure public health relevance rat Secp43 protein selenocysteine-tRNA selenocysteinyl-tRNA selenophosphate selenoprotein tRNA Ligase thiophosphate

项目摘要

DESCRIPTION (provided by applicant): Aminoacyl-tRNAs provide the interface between genetic information encoded in the DNA sequence of a gene and the amino acid sequence of the corresponding protein. The vast majority of organisms have been found to lack at least one canonical aminoacyl-tRNA synthetase. The corresponding amino acyl-tRNA is instead synthesized by either a non-canonical aminoacyl-tRNA synthetase or by an alternative pathway. A broad goal of the proposed work is to characterize structurally, biochemically and genetically enzymes and multi-enzyme and tRNA complexes involved in these new pathways for aminoacyl-tRNA synthesis that can then be used to uncover anti-infective targets for human pathogens and for unnatural amino acid incorporation. These general goals will be realized in four specific areas of the proposed work. (i) Selenocyste- inyl-tRNA formation is an essential process in humans, and likely in certain pathogens. Investigations of the enzymes involved, and of an alternative and still unknown route for Sec-tRNA formation will be of fundamental importance for understanding selenocysteine decoding and for exploiting this pathway as a way to combat human pathogens responsible for sleeping sickness and malaria. (ii) Using structure-based high-throughput random mutagenesis and computational design of pyrrolysyl-tRNA synthetase and tRNAPyl, genetic coding systems will be constructed for several lysine derivatives, which will be important for revealing the role of certain post-translational modifications in human and other eukaryotes. Translational quality control by the elongation factor EF-Tu will also be studied along side efforts to design elongation factor mutants that will specifically recognize the desired unnatural aminoacyl-tRNA. (iii) Several amino acids (selenocysteine, cysteine, glutamine, asparagine) are synthesized while attached to the tRNA. Complex formation, between the aminoacyl-tRNA synthetase, tRNA and biosynthetic enzyme(s)-possibly important for substrate channeling and protection of the translation system from mis-acylated tRNAs-will be investigated, to reveal how complex formation affects the enzymatic behavior of the component molecules. (iv) Our discovery and proposed characterization of a novel tRNA ligase(s) will lead to a better understanding of tRNA processing in humans and archaea. PUBLIC HEALTH RELEVANCE: The unexpected diversity of aminoacyl-tRNA synthesis (processes that maintain the coding relation- ship between DNA and protein) opens previously inaccessible frontiers in the biology of translation and post-translational protein modifications, the malfunction of which is linked to several human diseases including cancer, neurodegenerative and metabolic disorders. The proposed projects aim to characterize new routes for aminoacyl-tRNA formation and macromolecular complexes involved in aminoacyl-tRNA synthesis in order to uncover anti-infective targets for human pathogens (including the causative agents of sleeping sickness and malaria) and to design pathways for unnatural amino acid incorporation that will be applied to understanding the complex role of post-translational protein modifications in higher eukaryotes.

描述（由申请人提供）：氨基酰基trna提供基因DNA序列编码的遗传信息与相应蛋白质的氨基酸序列之间的接口。绝大多数生物体被发现至少缺乏一种典型的氨基酰基- trna合成酶。相应的氨基酰基- trna通过非规范的氨基酰基- trna合成酶或通过其他途径合成。所提出的工作的一个广泛目标是表征结构，生化和遗传酶，多酶和tRNA复合物参与这些新途径的氨基酰基-tRNA合成，然后可用于发现人类病原体的抗感染靶点和非天然氨基酸掺入。这些总体目标将在拟议工作的四个具体领域实现。(i) Selenocyste- inyl-tRNA的形成是人类必不可少的过程，可能在某些病原体中也是如此。对所涉及的酶以及另一种尚不清楚的Sec-tRNA形成途径的研究，对于理解硒代半胱氨酸解码和利用这一途径作为对抗导致昏睡病和疟疾的人类病原体的一种方式具有重要意义。（ii）利用基于结构的高通量随机突变和pyrolysyl - trna合成酶和tRNAPyl的计算设计，构建几种赖氨酸衍生物的遗传编码系统，这将对揭示某些翻译后修饰在人类和其他真核生物中的作用具有重要意义。此外，还将研究伸长因子EF-Tu的翻译质量控制，以及设计能够特异性识别所需的非天然氨基酰基trna的伸长因子突变体的努力。（iii）几种氨基酸（硒代半胱氨酸、半胱氨酸、谷氨酰胺、天冬酰胺）与tRNA结合合成。在氨基酰基-tRNA合成酶、tRNA和生物合成酶之间形成复合物——可能对底物通道和保护翻译系统免受误酰化tRNA的影响很重要——将被研究，以揭示复合物的形成如何影响组分分子的酶促行为。（iv）我们对一种新型tRNA连接酶的发现和提出的表征将有助于更好地理解人类和古细菌中tRNA的加工过程。