HPV DNA REPLICATION--NOVEL HOST FACTORS

候选人托马斯·梅伦迪获得了博士学位。1988年，来自加州大学洛杉矶分校，与Dan S. Ray. 在冷泉港实验室（1988-1994）担任博士后研究员期间，他与布鲁斯斯蒂尔曼博士一起研究猿病毒40（SV 40）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。 使用这个系统，他证明了牛PV DNA复制需要额外的细胞因子，而不是使用SV 40系统确定的那些。Melendy博士是微生物学（SUNY布法罗医学院）和细胞和分子生物学（Roswell Park癌症研究所）的助理教授，终身教职，也是研究生院的一员，可以接触来自各种项目的博士生。 生物医学研究大楼的研究环境，Melendy博士在那里拥有1,100平方米的办公室。英尺实验室和相邻的办公室，是优秀的，具有广泛的设施和设备。 每周举行研讨会和实验室间研究会议。Melendy博士目前得到NIH的研究支持（直到2002年），研究BPV DNA复制，并得到美国癌症协会的研究支持，研究DNA复制启动的机制。Melendy放弃在不久的将来进一步资助申请，并将允许他避免预计增加教学责任，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计划上。 长期职业目标是晋升为终身职位，保持对HPV DNA复制的积极研究计划。该研究项目的目标是研究病毒性性病的病原体HPV 11型的DNA复制。我们已经发现，像BPV一样，HPV 11需要额外的细胞DNA复制因子，而不是SV 40所需的那些。 HPV 11 DNA复制的重建将用于鉴定这些未鉴定的细胞因子。 HeLa细胞提取物缺乏HPV 11 DNA体外复制所需的单一细胞因子。 该因子与复溶反应所需的因子不同，并显著刺激复溶反应。 HeLa细胞提取物的互补作用将用于鉴定该细胞因子。将评估这些多种细胞因子在HPV 11 DNA复制过程中的作用，并鉴定这些蛋白质，如果未知，则克隆。

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.