ASSOCIATION OF TELOMERASE WITH TELOMERES IN HUMAN CELLS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 端粒对于染色体的稳定性是必不可少的。它们确保了对染色体末端的有效保护，并由一种专门的酶--端粒酶逆转录酶复制。端粒由TTAGGG序列的长重复序列(2-10kb)组成，末端有150-300个核苷酸长的TTAGGG单链突起。六种蛋白质复合体保护素与端粒特异结合，调节端粒的长度和复制，并确保端粒的保护。在表达端粒酶的细胞中，端粒酶的活动受到遮盖素的负调控，在表达该酶的细胞中，端粒的平均长度随着时间的推移保持稳定。这项规定涉及的事件尚不清楚。 这一提议的中心假设是端粒酶的招募受Shelterin复合体的调控。最初的目标将是初步开发和调整生化分析，以定量检测染色体末端的端粒酶(目标1)。这些分析将为分析其作用奠定基础。 酶在染色体末端的募集中的保护素(目标2)。将分析其他非保护素成分在端粒酶招募中的潜在积极作用(目标3)。 该提案的最终具体目标概述了PI的职业发展目标，以及将PI和实验室的工作纳入RCMI基因中心在亨特学院的目标的计划。 对端粒酶如何在其作用部位进行调控的理解在细胞水平上与癌症有关。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。在细胞分裂期间没有端粒维持的情况下，人类细胞最终停止分裂，这一过程被称为衰老，这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很大影响。

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.