STRUCTURE/FUNCTION STUDIES ON HUMAN TELOMERASE

这里提出的研究计划旨在让我利用自己在分子生物学和逆转录酶机制方面的博士后培训经验，结合我在突变方面的博士前教育，发展我在癌症研究方面的独立事业。计划中的项目将为我提供成功过渡到癌症研究的独特机会，使我能够利用我在艾滋病毒逆转录酶生物化学方面获得的丰富经验，并将其应用于端粒酶研究。部分培训经验将涉及与癌症研究领域公认的专家罗恩·德皮尼奥博士的合作。进一步的智力接触将来自与AECOM癌症中心调查人员的互动。AECOM的课程和研讨会将补充这些互动。最近，端粒酶逆转录酶(TERT)在启动转化和肿瘤发生过程中的关键作用已被确立。这使得它既是这些过程的核心参与者，也是癌症治疗干预的潜在靶点。因此，研究TERT的功能特性，了解其作用机制，确定影响其活性的因素，并勾画出对其活性至关重要的功能结构域和关键残基非常重要。这项拟议的研究将涉及对人类端粒酶的研究，以试图确定结构/功能关系。这将通过对重组的hTERT和天然hTERT的细胞提取物进行比较生化分析，通过定点突变鉴定hTERT中具有重要功能的残基，以及检查hTERT突变体的生物学活性来确定它们的生物学相关性来实现。

The research plan proposed here is designed to allow me to use my post-doctoral training experience in molecular biology and reverse transcriptase mechanism, in conjunction with my pre- doctoral education in mutagenesis, to develop my independent career in cancer research. The planned project will afford me the unique opportunity to make a successful transition into cancer research by allowing me to use the extensive experience I have acquired in HIV reverse transcriptase biochemistry and apply it to telomerase research. Part of the training experience will involve collaboration with Dr. Ron Depinho, an acknowledged expert in the field of cancer research. Further intellectual exposure will come from interactions with AECOM Cancer Center investigators. Courses and seminars at AECOM will complement these interactions. A key role for telomerase reverse transcriptase (TERT) has recently been established in events that initiate the processes of transformation and oncogenesis. This makes it is both a central player in these processes and a potential target for therapeutic intervention of cancer. Therefore, it is important to functionally characterize TERT, to learn its mechanism of action, identify factors that modify its activity and to delineate functional domains and key residues critical for its activity. The proposed research will involve the study of human telomerase in an attempt to determine structure/function relationships. This will be accomplished through comparative biochemical analysis between reconstituted hTERT and cellular extracts of native hTERT, identification of functionally important residues in hTERT via site-directed mutagenesis, and, examination of the biological activity of hTERT mutants to establish their biological relevance.