Analysis of telomerase reverse transcriptase

端粒酶逆转录酶分析

基本信息

批准号：
7939112
负责人：
NEAL F LUE
金额：
$ 24.55万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2009-09-30 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939112
关键词：
Apoptosis Architecture Binding Biochemical Genetics Biological Biological Assay Biological Models Biology Cells Chromatin Remodeling Factor Collection Complex DNA Data Development Electrostatics Enzymatic Biochemistry Enzymes Funding Goals Human In Vitro Maps Mediating Modeling Mutagenesis Mutation Names Neoplastic Cell Transformation Nucleic Acids Primer Extension Proteins Proteolysis RNA RNA Binding RNA Recognition Motif RNA-Protein Interaction Regulation Research Research Personnel Ribonucleoproteins Role Saccharomyces cerevisiae Saccharomycetales Sequence Alignment Series Site Structure Surface Telomerase Telomere Maintenance Telomere Shortening Terminal Repeat Sequences Tertiary Protein Structure Testing Yeasts base cancer cell cancer therapy clinical application crosslink genetic regulatory protein in vitro activity in vivo insight mutant neoplastic cell novel programs telomerase reverse transcriptase telomere

Apoptosis Architecture Binding Biochemical Genetics Biological Biological Assay Biological Models Biology Cells Chromatin Remodeling Factor Collection Complex DNA Data Development Electrostatics Enzymatic Biochemistry Enzymes Funding Goals Human In Vitro Maps Mediating Modeling Mutagenesis Mutation Names Neoplastic Cell Transformation Nucleic Acids Primer Extension Proteins Proteolysis RNA RNA Binding RNA Recognition Motif RNA-Protein Interaction Regulation Research Research Personnel Ribonucleoproteins Role Saccharomyces cerevisiae Saccharomycetales Sequence Alignment Series Site Structure Surface Telomerase Telomere Maintenance Telomere Shortening Terminal Repeat Sequences Tertiary Protein Structure Testing Yeasts base cancer cell cancer therapy clinical application crosslink genetic regulatory protein in vitro activity in vivo insight mutant neoplastic cell novel programs telomerase reverse transcriptase telomere

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项目摘要

DESCRIPTION (provided by applicant): Telomerase is a ribonucleoprotein complex responsible for replenishing the G-rich strand of the telomere terminal repeats. It employs a small, embedded segment of a stably associated RNA component as template, and pre-existing chromosomal ends as primers to synthesize telomere tracts. Telomerase activity is specifically activated in cancer cells, and promotes neoplastic transformation by endowing cells with unlimited replicative potential. Inhibition or depletion of telomerase in tumor cells leads to telomere shortening and apoptosis, validating the potential of anti-telomerase strategies in cancer therapy. The goal of this research is to understand telomerase mechanisms and regulation using the budding yeast Saccharomyces cerevisiae as a model system. Our studies during the past funding period have yielded a wealth of information on the structure and functions of the catalytic TERT protein. In addition, they revealed (i) two telomerase-specific domains within the catalytic TERT subunit that are responsible for a unique feature of telomerase enzymology, i.e., the ability to repetitively copy a short RNA template; (ii) an alpha-helical repeat domain (named TPR) in the Est1 p subunit that possesses a novel RNA-binding activity, and (iii) regulation of telomere structure and function by an ATP-dependent chromatin remodeling complex (the INO80 complex). Based on these findings, we propose a series of biochemical, genetic and cell biological studies directed toward (1) clarifying the mechanisms and biological significance of the protein- DNA and protein-RNA interactions mediated by telomerase components, and (2) ascertaining the roles of chromatin remodeling complexes in modulating telomere structures and telomerase activity. Results from the proposed studies are expected to broaden and deepen current understandings of telomerase mechanisms and provide insights into a novel aspect of telomere regulation. Because almost all of the factors that will be examined are conserved between yeast and humans, we anticipate that many of the findings will be applicable to human telomeres and telomerase, and will facilitate the development of clinical applications based on telomere biology.

描述（由申请人提供）：端粒酶是一种核糖核蛋白复合物，负责补充端粒末端重复量富含G的链。它采用稳定关联的RNA分量的小段作为模板，并预先存在的染色体末端作为合成端粒区域的底漆。端粒酶活性在癌细胞中特异性激活，并通过赋予无限复制潜力的细胞来促进肿瘤转化。肿瘤细胞中端粒酶的抑制或耗竭会导致端粒缩短和凋亡，从而验证了抗细胞瘤策略在癌症治疗中的潜力。这项研究的目的是使用酿酒酵母作为模型系统了解端粒酶机制和调节。在过去的资金期间，我们的研究产生了有关催化TERT蛋白的结构和功能的大量信息。此外，他们揭示了（i）催化TERT亚基内两个端粒酶特异性域，这些结构域负责端粒酶酶的独特特征，即重复复制短RNA模板的能力；（ii）EST1 P亚基中具有新型RNA结合活性的α-螺旋重复域（命名为TPR），以及（iii）通过ATP依赖性染色质重塑复合物（INO80复合物）对端粒结构和功能进行调节。基于这些发现，我们提出了一系列针对（1）阐明蛋白质DNA和蛋白质-RNA相互作用的机制和生物学意义的生化，遗传和细胞生物学研究，以及（2）确定染色质重塑型在模块化望远镜结构和触发型中的作用。拟议研究的结果预计将扩大和加深对端粒酶机制的理解，并为端粒调节的新方面提供见解。因为几乎所有将要检查的因素都是在酵母和人类之间保存的，所以我们预计许多发现将适用于人类端粒和端粒酶，并将促进基于端粒生物学的临床应用的开发。