Structural Study of Vertebrate Telomerase RNA

脊椎动物端粒酶RNA的结构研究

• 批准号: 8784637
• 负责人: Yaqiang Wang
• 金额: $ 5.33万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784637
• 关键词: Acylation; Affinity; Antineoplastic Agents; Apoptosis; Architecture; Binding; Binding Sites; Biochemical; Biological Assay; Biological Process; Boundary Elements; C-terminal; Calorimetry; Catalytic Domain; Cell Aging; Cells; Chemicals; Chromosomes; Complex; DNA; DNA Repair; DNA biosynthesis; Data; Drug Design; Drug Targeting; Elements; Ensure; Fellowship; Fishes; Future; Goals; Hepatitis C virus; Human; Hydroxyl Radical; Internal Ribosome Entry Site; Japanese Killifish; Length; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Modeling; Molecular; N-terminal; NMR Spectroscopy; Normal Cell; Nuclear Magnetic Resonance; Primer Extension; RNA; RNA Recognition Motif; RNA Sequences; RNA-Directed DNA Polymerase; Resolution; Ribonucleoproteins; Roentgen Rays; Role; Sampling; Shapes; Solutions; Structural Models; Structure; Telomerase; Telomerase RNA Component; Telomere Shortening; Testing; Titrations; Virus Replication; X-Ray Crystallography; base; cancer cell; in vitro activity; insight; prevent; protein complex; public health relevance; research study; senescence; small molecule; telomerase reverse transcriptase; telomere; tool

DESCRIPTION (provided by applicant): Telomerase is the ribonucleoprotein complex that catalyzes the addition of telomere DNA repeats onto the 3' ends of linear chromosomes. Telomeres protect the ends of chromosomes from being recognized by the cellular DNA repair machinery as double strand breaks and also prevent end joining. In the absence of active telomerase, telomeres shorten with each round of DNA replication. Once a critical amount of telomere is lost, the cell enters replicative senescence and sometimes undergoes apoptosis. The high level of telomerase activity present in most cancers allows this roadblock to be avoided, making telomerase a potential drug target. Telomerase RNA and telomerase reverse transcriptase comprise the essential elements for telomerase activity in vitro. The goal of this fellowship will be to solve the high- resolution structure of medaka telomerase RNA P2ab in the catalytic core by NMR, investigate a potential of human telomerase RNA P2ab as a drug target to inhibit telomerase activity, and obtain small angle X- ray scattering structures of the medaka telomerase RNA both free and in complex with the telomerase reverse transcriptase. The specific aims are to (1) investigate the structure, dynamics, and function of medaka telomerase RNA P2ab domain, (2) investigate human telomerase RNA J2a/b as a drug target to inhibit telomerase activity, (3) model medaka telomerase RNA using NMR and small angle X-ray scattering, (4) probe the structure and interactions between the medaka telomerase RNA and domains of the telomerase reverse transcriptase using small angle X-ray scattering and X-ray crystallography. These experiments will reveal the structure of the medaka telomerase RNA catalytic core and the intermolecular contacts between the RNA and the subdomain of the telomerase reverse transcriptase.

描述（由申请方提供）：端粒酶是一种核糖核蛋白复合物，催化端粒DNA重复序列添加到线性染色体的3'端。端粒保护染色体的末端不被细胞DNA修复机制识别为双链断裂，并且还防止末端连接。在没有活性端粒酶的情况下，端粒随着每一轮DNA复制而缩短。一旦失去了临界量的端粒，细胞就进入复制性衰老，有时会发生凋亡。大多数癌症中存在的高水平端粒酶活性可以避免这一障碍，使端粒酶成为潜在的药物靶点。端粒酶RNA和端粒酶逆转录酶是端粒酶在体外发挥活性的重要组成部分。本研究的目标是通过NMR解析青鳉端粒酶RNA P2 ab在催化核心的高分辨率结构，研究人端粒酶RNA P2 ab作为抑制端粒酶活性的药物靶点的潜力，并获得青鳉端粒酶RNA游离和与端粒酶逆转录酶复合的小角X射线散射结构.本研究的具体目标是：（1）研究青鳉鱼端粒酶RNA P2 ab结构域的结构、动力学和功能;（2）研究人端粒酶RNA J2 a/B作为抑制端粒酶活性的药物靶点;（3）利用NMR和小角X射线散射技术模拟青鳉鱼端粒酶RNA，（4）利用小角X射线散射和X-射线光散射技术研究青鳉端粒酶RNA的结构和与端粒酶逆转录酶结构域之间的相互作用。射线晶体学这些实验将揭示青鳉端粒酶RNA催化核心的结构和RNA与端粒酶逆转录酶亚结构域之间的分子间接触。