Telomerase function and regulation in a new model system

新模型系统中的端粒酶功能和调节

• 批准号: 7094239
• 负责人: NEAL F LUE
• 金额: $ 29.2万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7094239
• 关键词: Candida albicans; enzyme activity; enzyme mechanism; fungal genetics; genetic recombination; immunoprecipitation; intermolecular interaction; mutant; protein protein interaction; protein purification; protein structure function; site directed mutagenesis; telomerase; telomere; Candida albicans; enzyme activity; enzyme mechanism; fungal genetics; genetic recombination; immunoprecipitation; intermolecular interaction; mutant; protein protein interaction; protein purification; protein structure function; site directed mutagenesis; telomerase; telomere

DESCRIPTION (provided by applicant): Telomerase is a ribonucleoprotein complex responsible for extending the dG-rich strand of the telomere terminal repeats. It employs a small segment of a stably associated RNA component as template, and pre-existing chromosomal ends as primers to accomplish reverse transcription. Telomerase activity isspecifically activated in cancer cells, and promotes neoplastic transformation by endowing cells with unlimited replicative potential. Selective inactivation of telomerase in tumor cells leads to telomere shortening and apoptosis, validating the potential of telomerase inhibitors in anti-cancer therapy. The goal of this research is to investigate telomerase regulation and function in Candida albicans, a relatively unexplored experimental system. In contrast to other well-studied fungi, Candida possesses longer telomere tracts, a regular extended telomere repeat, and exhibits more active recombination at telomeres. Because of these unusual features, we have been able to develop unique biochemical and molecular toolsfor addressing fundamental issues in the field. Most importantly, we have been able to demonstrate for the first time the effects of two regulatory factors on telomerase activity in vitro, and are thus in a position to approach telomerase regulation using the powerful combination of yeast genetics and biochemistry. Our preliminary studies suggest two major hypotheses: (1) the CaEstlp and CaEst3p subunit of the telomerase complex can activate telomerase function through protein-protein and/or protein-DNA interaction, and (2) some telomerase subunits can participate in the protection of telemere ends independent of their telomere extension function. To examine these hypotheses, we will utilize a series of biochemical, genetic, and cell biological assays to analyze Candida telomeres and telomerase inappropriate mutant strains. We will also develop purified in vitro systems to assess telomerase activation by regulatory factors, and to investigate relevant protein-DNA and protein-protein interactions. We anticipate that our findings will contribute to a detailed molecular understanding of telomerase function and regulation.

描述（由申请人提供）：端粒酶是一种核糖核蛋白复合物，负责扩展端粒末端重复的富含DG的链。它使用稳定关联的RNA成分的一小段作为模板，并预先存在染色体末端作为完成逆转录的底漆。在癌细胞中特别激活的端粒酶活性，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选择性失活会导致端粒缩短和凋亡，从而验证端粒酶抑制剂在抗癌治疗中的潜力。 这项研究的目的是研究念珠菌白色念珠菌的端粒酶调节和功能，这是一个相对未开发的实验系统。与其他研究的真菌相比，念珠菌具有更长的端粒段，常规的端粒重复延伸，并在端粒中表现出更积极的重组。由于这些不寻常的特征，我们已经能够开发出独特的生化和分子工具来解决该领域的基本问题。最重要的是，我们能够首次证明两个调节因素对体外端粒酶活性的影响，因此可以使用酵母遗传学和生物化学的强大组合来接近端粒酶调节。我们的初步研究提出了两个主要的假设：（1）端粒酶复合物的CAESTLP和CAESTLP和CAEST3P亚基可以通过蛋白质蛋白质和/或蛋白质DNA相互作用激活端粒酶功能，（2）一些端粒酶亚基可以参与远程远程远程远程远程远程远程远程分析功能的保护。为了审查这些假设，我们将利用一系列的生化，遗传和细胞生物学测定法来分析念珠菌端粒和端粒酶不适当的突变株。我们还将开发出纯化的体外系统，以评估调节因素的端粒酶激活，并研究相关的蛋白-DNA和蛋白质 - 蛋白质相互作用。我们预计我们的发现将有助于对端粒酶功能和调节的详细分子理解。