CRYSTAL STRUCTURE DETERMINATION OF QUEOSINE-SYNTHESIS AND OTHER TRNA MODIFICATIO

奎辛合成和其他 TRNA 修饰的晶体结构测定

• 批准号: 7954428
• 负责人: MANAL A SWAIRJO
• 金额: $ 0.21万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954428
• 关键词: Anabolism; Antibiotics; Anticodon; Automation; Biochemical; Cells; Codon Nucleotides; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Data; Development; Drug Delivery Systems; Enzymes; Funding; Genomics; Grant; HIV; Institution; Ligands; Methods; Modification; Pathway interactions; Pharmacologic Substance; Prokaryotic Cells; Research; Research Personnel; Resources; Ribosomes; Shigella Infections; Source; Structure; Technology; Time; Transfer RNA; Translations; United States National Institutes of Health; analog; beamline; comparative; structural biology; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have used SSRL beam time on this proposal to conduct crystallographic studies on enzymes responsible for the biosynthesis of two tRNA modifications: queosine (Q) and threonyl carbamoyladenosine (t6A). These are two modifications of the tRNA anticodon loop that alter codon definitions and fine tune ribosome function during translation. Q biosynthesis is an established drug target for anti-shigellosis antibiotics. Similarly, due to its requirement for HIV replication, t6A modification of host-cell tRNA has been proposed as a potential target for the development of anti-HIV pharmaceuticals. Using a combination of comparative genomic, biochemical, and structural methods, we identified enzymes involved in the biosynthesis of Q and t6A. Using SSRL x-rays, we determined the structure of GCYH-IB, our newly discovered and first enzyme in the Q biosynthesis pathway in prokaryotes, in complex with a substrate analog and we collected native and heavy-atom derivative data for YkvM, the enzyme that catalyzes the third step in Q biosynthesis. We also determined the crystal structure of YrdC, a universally conserved and essential enzyme involved in t6A biosynthesis, in complex with its ATP substrate. In this renewal, we plan to start crystallographic studies of GCYH-IBinhibitor complexes and of QueA, which catalyzes a late step in Q biosynthesis, in complex with its tRNA substrate. We currently have crystals of GCYH-IB-ligand complexes and crystallization of the latter complex is underway. We plan to continue to use the SSRL beamline automation technology and remote access for these studies.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们已经在该提案上使用了SSRL束时间，以对负责两种tRNA修饰的生物合成的酶进行晶体学研究：queosine（q）和三酰胺甲bamoyladenosine（T6A）。这是TRNA反密码子循环的两个修饰，它们在翻译过程中改变了密码子的定义和微调核糖体功能。 Q生物合成是抗硫醇抗生素的既定药物。同样，由于其对HIV复制的要求，T6A对宿主细胞tRNA的修饰是抗HIV药物开发的潜在靶标。使用比较基因组，生化和结构方法的组合，我们确定了参与Q和T6A生物合成的酶。使用SSRL X射线，我们确定了GCYH-IB的结构，我们在原核生物的Q生物合成途径中新发现和第一种酶与底物类似物复杂，我们收集了YKVM的本地和重原子数据，YKVM，YKVM，YKVM，该酶是Quosynsys in Q Biosyntysessissys的第三步的酶。我们还确定了YRDC的晶体结构，这是一种与T6A生物合成有关的普遍保守和必需的酶，与其ATP底物复杂化。在此续约中，我们计划开始对GCYH-抑制剂复合物和QueA的晶体学研究，该研究与其TRNA底物相结合，从而促进了Q生物合成的后期一步。目前，我们有GCYH-IB-配体配合物的晶体，后者复合物的结晶正在进行中。我们计划继续为这些研究使用SSRL梁线自动化技术和远程访问。