HPV DNA REPLICATION: NOVEL HOST FACTORS

候选人托马斯·梅伦迪（Thomas Melendy）于1988年在加州大学洛杉矶分校获得博士学位，与丹·s·雷（Dan S. Ray）博士一起研究锥虫体DNA拓扑异构酶。1988-1994年在冷泉港实验室做博士后时，与Bruce Stillman博士一起研究类人猿病毒40 （SV40）的DNA复制因子和机制。Melendy博士随后开发了一种部分纯化的体外牛乳头瘤病毒（PV） DNA复制系统。使用该系统，他证明牛PV DNA复制需要额外的细胞因子，而不是使用SV40系统鉴定的那些。Melendy博士是微生物学（纽约州立大学布法罗医学院）和细胞与分子生物学（罗斯威尔公园癌症研究所）的助理教授，终身教授，也是研究生院的一员，可以接触来自不同项目的博士生。生物医学研究大楼的研究环境，梅伦迪博士在那里有1100平方英尺。实验室和相邻的办公室，是优秀的，拥有各种设施和设备。每周举行研讨会和实验室间研究会议。Melendy博士目前获得美国国立卫生研究院（NIH）的研究支持（直到2002年），研究BPV DNA复制，并获得美国癌症协会的研究支持，研究DNA复制起始机制。K02奖的支持将使Melendy博士在不久的将来放弃进一步的资助申请，并使他能够避免预计的教学责任增加，使他能够在职业发展的重要时期将90%的精力集中在他的研究项目上。长期的职业目标是晋升到终身职位，保持对HPV DNA复制的积极研究计划。本研究项目的目的是研究HPV 11型的DNA复制，HPV 11型是一种病毒性STD的病原。我们发现，与BPV一样，HPV 11需要额外的细胞DNA复制因子，而不是SV40所需的。HPV 11 DNA复制的重构将用于识别这些未识别的细胞因子。HeLa细胞提取物缺乏体外HPV 11 DNA复制所需的单一细胞因子。该因子与重构反应所需的因子不同，能显著地促进重构反应。HeLa细胞提取物的补充将用于鉴定这种细胞因子。这些多种细胞因子在HPV 11 DNA复制过程中的作用将被评估，这些蛋白质将被识别，如果未知，将被克隆。

The candidate, Thomas Melendy, earned his Ph.D. from UCLA in 1988, studying trypanosomal DNA topoisomerases with Dr. Dan S. Ray. While a postdoctoral fellow at Cold Spring Harbor Laboratory (1988-1994), he worked with Dr. Bruce Stillman studying simian virus 40 (SV40) DNA replication factors and mechanisms. Dr. Melendy then developed a partially purified in vitro bovine papillomavirus (PV) DNA replication system. Using this system he demonstrated that bovine PV DNA replication requires additional cellular factors other than those identified using the SV40 system. Dr. Melendy is an Assistant Professor of Microbiology (SUNY Buffalo School of Medicine) and Cellular and Molecular Biology (Roswell Park Cancer Institute), tenure-track, and a member of the Graduate Faculty with access to PhD students from various programs. The research environment in the Biomedical Research Building, where Dr. Melendy has a 1,100 sq. ft. laboratory and adjacent office, is excellent, with a wide range of facilities and equipment. Seminars and inter-laboratory research meetings are held weekly. Dr. Melendy currently has research support from the NIH (until 2002) to study BPV DNA replication, and from the American Cancer Society to study mechanisms of DNA replication initiation. Support from a K02 award will allow Dr. Melendy to forego further grant applications for the near future and will permit him to avoid projected increases in teaching responsibilities, allowing him to focus over 90 percent of his efforts on his research program at an important time in his career development. Long term career goals are to advance to a tenured position, maintaining an active research program on HPV DNA replication. The goals for this research project are to study the DNA replication of HPV type 11, the etiological agent of a viral STD. We have found that like BPV, HPV 11 requires additional cellular DNA replication factors beyond those required by SV40. The reconstitution of HPV 11 DNA replication will be used to identify these unidentified cellular factors. Extracts from HeLa cells are deficient in a single cellular factor required for HPV 11 DNA replication in vitro. This factor is unique from those required for the reconstituted reaction and dramatically stimulates the reconstituted reaction. The complementation of HeLa cell extracts will be used to identify this cellular factor. The role of these multiple cellular factors in the HPV 11 DNA replication process will be evaluated and these proteins will be identified, and if unknown, cloned.