STRUCTURE/FUNCTION STUDIES ON HUMAN TELOMERASE

这里提出的研究计划是为了让我利用我在分子生物学和逆转录酶机制方面的博士后培训经验，结合我在诱变方面的博士前教育，在癌症研究方面发展我的独立事业。计划中的项目将为我提供一个独特的机会，使我能够利用我在HIV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，从而成功过渡到癌症研究领域。部分培训经验将涉及与Ron Depinho博士合作，他是癌症研究领域公认的专家。进一步的智力暴露将来自与AECOM癌症中心研究人员的互动。AECOM的课程和研讨会将补充这些互动。端粒酶逆转录酶（TERT）最近在启动转化和肿瘤发生过程的事件中发挥了关键作用。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，重要的是功能表征TERT，了解其作用机制，确定改变其活性的因素，并描绘其活性的功能域和关键残基。拟议的研究将涉及对人类端粒酶的研究，试图确定结构/功能关系。这将通过对重组hTERT和天然hTERT的细胞提取物进行比较生化分析，通过定点诱变鉴定hTERT中功能重要的残基，以及检查hTERT突变体的生物活性以确定其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.