The role of lysine acetylation of human threonyl-tRNA synthetase

人苏氨酰-tRNA合成酶赖氨酸乙酰化的作用

• 批准号: 10112444
• 负责人: Chenguang Fan
• 金额: $ 45万
• 依托单位: UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112444
• 关键词: 5' Untranslated Regions; Acetylation; Address; Affect; Amino Acyl-tRNA Synthetases; Aminoacylation; Anticodon; Bacteria; Binding; Binding Proteins; Biochemical; Cells; Codon Nucleotides; Crystallization; Development; Diagnostic; Drug Targeting; Enzymes; Escherichia coli; Family; Functional disorder; Genetic Code; Genetic Transcription; Glutamine; Goals; Growth; Homologous Gene; Human; Human Cell Line; Impairment; Laboratories; Link; Lysine; Malignant Neoplasms; Medical; Methods; Molecular; Mutation; Neuropathy; Oncogenic; Play; Protein Biosynthesis; Proteins; Proteomics; RNA Splicing; Research; Role; Series; Serine; Site; Solid; Somatic Mutation; Structure; Students; System; Testing; Therapeutic; Threonine; Threonine-Specific tRNA; Threonine-tRNA Ligase; Transcript; Transfer RNA Aminoacylation; Translations; Tumor Suppressor Proteins; Underrepresented Populations; Variant; cancer cell; cancer diagnosis; cancer therapy; cancer type; cell injury; drug development; experience; human disease; innovation; insight; novel; novel strategies; programs; targeted treatment; therapeutic development; therapeutic target; undergraduate student

PROJECT SUMMARY Aminoacyl-tRNA synthetases (AARSs) are a family of essential enzymes for protein synthesis, which also play important roles in regulating transcription, translation, and RNA splicing. Somatic mutations of AARSs have been found in many human diseases, making them favorable targets for drug development and medical therapies. Besides mutation, proteomic studies have also identified a series of acetylated lysine residues in AARSs from cancer cells but without further studies. So there is a critical need to identify the association of AARS acetylation with cancer formation. The overall goal of this project is to study the role of lysine acetylation of one AARS, threonyl-tRNA synthetase (ThrRS) in human. Given that the homologue of ThrRS in bacteria has been shown to be regulated by acetylation, the hypothesis is that lysine acetylation affects human ThrRS functions which facilitates cancer formation. To test this hypothesis, three specific aims are proposed. Aim 1 is to identify the site- and cancer-specific effects of lysine acetylation on ThrRS functions biochemically. Aim 2 is to demonstrate the impacts of acetylation on ThrRS structurally. Aim 3 is to identify the impacts of ThrRS acetylation on global translation and translation of cancer-associated proteins, i.e. oncogenic proteins and tumor suppressor in human cell lines. Oncogenic transformation tests on normal human cells expressing acetylated ThrRS variants will be performed to find whether ThrRS acetylation is an initiator for cancer formation or a cellular adaption for cancer growth. This proposal is innovative because: First, it will be the first to study lysine acetylation of human ThrRS, and is expected to identify novel mechanisms for ThrRS-associated cancer formation. Second, to address the problem that the classic glutamine-substitution method for acetylation studies is not always effective, the genetic code expansion approach will be applied in this proposal to co-translationally incorporate acetyllysine at controlled sites in order to produce site-specifically acetylated ThrRS variants which have been identified in cancer cells. The proposed research is significant because it will provide solid evidence for the association of ThrRS acetylation with cancer formation. Ultimately, the proposed studies on ThrRS acetylation are expected to identify novel targets for cancer diagnosis and treatment. Specific for the AREA program, the research team for this project will be composed primarily of undergraduate students including those from underrepresented groups. Both well-established methods and new approaches will be applied in this proposal to make the project promising, and in the meantime, to strengthen students’ research experiences.

项目摘要 氨基酰基-TRNA合成酶（AARSS）是蛋白质合成的必需酶家族，也可以发挥作用 在调节转录，翻译和RNA剪接方面的重要作用。 AARS的体细胞突变已经 在许多人类疾病中发现，使其成为药物开发和医疗疗法的有利目标。 除突变外，蛋白质组学研究还确定了来自AARS中的一系列乙酰化赖氨酸残基 癌细胞，但没有进一步的研究。因此，迫切需要确定AARS乙酰化的关联 与癌症形成。该项目的总体目标是研究一个AARS的赖氨酸乙酰化的作用， 人类中的threonyl-tRNA合成酶（THRRS）。鉴于已经显示了细菌中THRR的同源物 要受乙酰化调节，假设是赖氨酸乙酰化会影响人类THRR的功能 促进癌症形成。为了检验这一假设，提出了三个具体目标。目标1是确定 赖氨酸乙酰化对THRR的位点和癌症特异性作用在生化上起作用。目标2是证明 乙酰化对THRR的影响在结构上。 AIM 3是确定THRRS乙酰化对全球的影响 癌症相关蛋白的翻译和翻译，即人类的致癌蛋白和肿瘤抑制剂 细胞系。对表达乙酰化THRR的正常人细胞上的致癌转化测试将是 进行的是发现THRRS乙酰化是癌症形成还是癌症的细胞适应性的引发剂 生长。该提议具有创新性，因为：首先，它将是第一个研究人类THRR的赖氨酸乙酰化， 并有望确定与THRS相关癌症形成的新型机制。第二，解决 乙酰化研究的经典谷氨酰胺固定方法并不总是有效的问题 该提案将采用代码扩展方法，以共同掺入乙酰透析 受控位点以产生特定于位置的乙酰化的THRRS变体，并已在 癌细胞。拟议的研究很重要，因为它将为关联提供可靠的证据 与癌症形成的THRR乙酰化。最终，关于THRRS乙酰化的拟议研究有望 确定癌症诊断和治疗的新目标。特定于该地区计划的研究团队 该项目将主要由本科生组成，包括来自代表性不足的小组的学生。 在本提案中，将采用公认的方法和新方法，以使项目承诺， 同时，增强学生的研究经验。