TRANSCRIPTIONAL REGULATION OF HUMAN 17-HYDROXYLASE GENE

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In accordance with the Physician Scientist Award (PSA), the candidate and the sponsor, in conjunction with the Dept of Obstetrics and Gynecology and the Graduate Program in Endocrinology at UCSF, have constructed a detailed 2-phase 5-year training program intended to provide the candidate with an intensive research experience. This program is an integral part of her candidacy for a Ph.D. in Endocrinology. Phase I involves fully registered enrollment in eight graduate division courses in 6 departments, regular attendance at various seminars and three lab rotations. These will be done concurrently with the candidate's training as a fellow in Reproductive Endocrinology in the Dept. of Obstetrics and Gynecology. This experience will fulfill the Phase I requirements of the PSA and fulfill the requirements to advance to candidacy in the Ph.D. program. Phase II is 100% lab research under the direct supervision of the primary sponsor. This research will also constitute the applicants Ph.D. thesis. During phase II, the candidate's research effort will focus on the transcriptional regulation of the human P450c17 gene, the interaction between specific cis- elements and trans-acting factors which leads to the tissue-specific aims: (1) Investigate the potential regulatory role of the intragenic sequences on hormonally regulated transcription by stable transfection. Thus far only the 5' flanking region has been studied for hormone-responsiveness. However, in some other systems, intragenic sequences have been shown to modulate transcription. (2) Define the specific cis-elements involved in tissue-specificity and hormone-responsiveness. Transient transfection experiments will be performed with plasmids containing fragments of the 5'flanking region linked to a heterologous promotor and CAT reporter gene. (3) Establish a murine cell line which expresses P450c17 endogenously. Study of the P450c17 gene has been limited by the lack of a cell line with endogenous P450c17 expression. We will construct transgenic mice with P450c17 targeted SV40 T-antigen expression. This approach has been used with success by collaborators at this institution to establish differentiated cell lines. (4) Redefine cis-elements involved in tissue- specificity and hormone-responsiveness by transient transfection with cell lines established in aim #3. Interaction between specific nuclear factors and these cis-elements will be investigated. Successful completion of these aims will add substantially to our understanding of the hormonally regulated tissue-specific expression of this key enzyme in steroidogenesis. Successful completion of this training program will equip the candidate to initiate an independent research career studying the molecular underpinnings of human reproduction.