HPV DNA REPLICATION: NOVEL HOST FACTORS

候选人托马斯·梅伦迪于1988年在加州大学洛杉矶分校获得博士学位，与丹·S·雷博士一起研究锥虫DNA拓扑异构酶。作为冷泉港实验室的博士后研究员(1988-1994)，他与Bruce Stillman博士一起研究猿猴病毒40(SV40)DNA复制因素和机制。梅伦迪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。利用这个系统，他证明了牛PV DNA复制需要额外的细胞因素，而不是使用SV40系统所确定的那些因素。Melendy博士是纽约州立大学布法罗医学院微生物学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，终身教职，也是研究生院的成员，可以接触到不同项目的博士生。生物医学研究大楼的研究环境，梅伦迪博士在那里有1,100平方英尺。英国《金融时报》实验室与办公楼毗邻，设备齐全，设施齐全。每周举行研讨会和实验室间研究会议。梅伦迪博士目前得到了美国国立卫生研究院(NIH)研究BPV DNA复制的研究资助(直到2002年)，并得到了美国癌症协会的支持，研究DNA复制启动的机制。K02奖项的支持将使梅伦迪博士在不久的将来放弃进一步的拨款申请，并将使他避免预期的教学责任增加，使他能够在职业发展的重要时刻将90%以上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。这项研究的目标是研究HPV11型的DNA复制，HPV11型是一种病毒性STD的病原体。我们发现，与BPV一样，HPV11需要额外的细胞DNA复制因子，而不是SV40所要求的因子。HPV11DNA复制的重组将用于鉴定这些未知的细胞因素。HeLa细胞提取物缺乏HPV11DNA在体外复制所需的单一细胞因子。该因子不同于重组反应所需的那些，并且极大地刺激了重组反应。HeLa细胞提取液的互补性将用于鉴定该细胞因子。将评估这些多种细胞因素在HPV11DNA复制过程中的作用，并将鉴定这些蛋白质，如果未知，还将进行克隆。

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.