ASSOCIATION OF TELOMERASE WITH TELOMERES IN HUMAN CELLS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 端粒对染色体稳定性至关重要。它们确保有效地保护染色体末端，并通过一种专用的酶--端粒酶逆转录酶进行复制。端粒由TTAGGG序列的长（2- 10 kb）重复序列组成，末端为150 -300个核苷酸长的TTAGGG单链突出端。六种蛋白复合物shelterin特异性结合端粒，调节其长度和复制，并确保其保护。在表达端粒酶的细胞中，端粒酶的作用受到shelterin的负调控，导致端粒平均长度随时间推移而稳定。该法规涉及的事件尚不清楚。 这个提议的中心假设是端粒酶的募集受shelterin复合物的调节。最初的目标将是初步开发和适应生化测定，以定量检测染色体末端的端粒酶（目标1）。这些试验将为分析其作用奠定基础 shelterin在招募酶染色体末端（目的2）。将分析其他非shelterin组分在端粒酶募集中的潜在积极作用（目的3）。 本提案的最终具体目标概述了PI职业发展的目标，以及将PI和实验室的努力纳入亨特学院RCMI基因中心目标的计划。 对端粒酶在其作用位点如何调节的理解与细胞水平的癌症有关。所有癌细胞最终都激活了端粒维持机制，为它们提供了无限的复制潜力。在细胞分裂过程中缺乏端粒维持的情况下，人类细胞最终会停止分裂，这一过程被称为衰老，衰老是一种重要的肿瘤抑制因子 机制因此，拟议的工作将对人类健康产生重大影响。

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. Telomeres are essential for chromosome stability. They ensure effective protection of chromosome ends, and are replicated by a dedicated enzyme, the telomerase reverse transcriptase. Telomeres consist of long (2-10kb) repeats of the TTAGGG sequence and end with a150-300 nucleotides-long TTAGGG single stranded overhang. The six protein complex shelterin specifically binds to telomeres, regulates their length and replication, and ensures their protection. The action of telomerase is negatively regulated by shelterin, accounting for stable average telomere length over time, in cells that express the enzyme. The events involved in this regulation are unclear. The central hypothesis of this proposal is that the recruitment of telomerase is regulated by the shelterin complex. The initial objective will be to initially develop and adapt biochemical assay to quantitatively detect telomerase at chromosome ends (Aim 1). These assays will lay the foundation for the analysis of the role of shelterin in the recruitment of the enzyme to chromosome ends (Aim 2). Other non-shelterin components will be analyzed for their potential positive role in telomerase recruitment (Aim 3). The final specific aim of this proposal outlines the goals for the PI's career development, as well as the plan for the inclusion of the PI and laboratory's efforts into the RCMI Gene Center's goals at Hunter College. The understanding of how telomerase is regulated at its site of action is relevant to cancer at the cellular level. All cancer cells have ultimately activated a mechanism of telomere maintenance with provides them with infinite replicative potential. In the absence of telomere maintenance during cell division, human cells eventually cease to divide, a process called senescence, which is an important tumor suppressor mechanism. Therefore, the proposed work will have high impact on human health.