HPV DNA REPLICATION--NOVEL HOST FACTORS

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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.