Structural Study of Vertebrate Telomerase RNA

脊椎动物端粒酶RNA的结构研究

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：端粒酶是一种核糖核蛋白复合物，可催化将端粒 DNA 重复序列添加到线性染色体的 3' 末端。端粒可保护染色体末端不被细胞 DNA 修复机制识别为双链断裂，并防止末端连接。在缺乏活性端粒酶的情况下，端粒会随着每轮 DNA 复制而缩短。一旦丢失了临界数量的端粒，细胞就会进入复制衰老，有时甚至会发生细胞凋亡。大多数癌症中存在高水平的端粒酶活性，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和端粒酶逆转录酶构成了体外端粒酶活性的基本要素。该奖学金的目标是通过核磁共振解析青鳉端粒酶RNA P2ab催化核心的高分辨率结构，研究人端粒酶RNA P2ab作为抑制端粒酶活性的药物靶点的潜力，并获得小角度X射线青鳉端粒酶RNA的散射结构，包括游离的和与端粒酶逆转录酶复合的。具体目标是 (1) 研究青鳉端粒酶 RNA P2ab 结构域的结构、动力学和功能，(2) 研究人端粒酶 RNA J2a/b 作为抑制端粒酶活性的药物靶点，(3) 使用青鳉端粒酶 RNA 模型NMR和小角X射线散射，(4)利用小角X射线散射探测青鳉端粒酶RNA与端粒酶逆转录酶结构域之间的结构和相互作用， X射线晶体学。这些实验将揭示青鳉端粒酶RNA催化核心的结构以及RNA与端粒酶逆转录酶亚结构域之间的分子间接触。