TRANSCRIPTIONAL REGULATION OF HUMAN 17-HYDROXYLASE GENE

人17-羟化酶基因的转录调控

• 批准号: 3086599
• 负责人: WEN-HUI SHEN
• 金额: $ 7.72万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-06-01 至 1997-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3086599
• 关键词: genetic regulation; genetic regulatory element; genetic transcription; genetically modified animals; hormone regulation /control mechanism; human genetic material tag; human tissue; laboratory mouse; steroid 17alpha monooxygenase; steroid hormone; steroid hormone biosynthesis; tissue /cell culture; transcription factor; transfection

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.

根据医师科学家奖（PSA），候选人和 申办方与妇产科部门合作， 加州大学旧金山分校内分泌学研究生课程，已经建立了一个详细的 2-为期5年的培训计划，旨在为候选人提供 密集的研究经验。 这个项目是她 博士候选人资格内分泌学 第一阶段涉及全面注册 在6个系的8个研究生部课程中注册，定期 参加各种研讨会和三次实验室轮换。 这些都将完成 同时，候选人的培训作为一个研究员在生殖 内分泌科妇产科 这段经历 将满足PSA第一阶段的要求，并满足 进入博士候选人资格的要求程序. 二期 在主要申办者的直接监督下进行100%的实验室研究。 这项研究也将构成申请人博士学位。论文 期间 第二阶段，候选人的研究工作将集中在转录 人P450 c17基因的调控，特异性顺式- 导致组织特异性目标的要素和反式作用因子： (1)研究基因内序列的潜在调控作用 通过稳定转染来调节转录。 迄今 仅研究了5 ′侧翼区的酶切反应性。 然而，在一些其他系统中，基因内序列已被证明是 调节转录。 (2)定义参与的特定顺式元件 组织特异性和免疫反应性。 瞬时转染 实验将用含有该基因片段的质粒进行。 5 '侧翼区连接到异源启动子和CAT报告基因。 (3)建立内源性表达P450 c17的小鼠细胞系。 对P450 c17基因的研究受到缺乏具有以下特征的细胞系的限制： 内源性P450 c17表达。 我们将构建转基因小鼠， P450 c17靶向SV 40 T抗原表达。 这一方法已被用于 通过这个机构的合作者成功地建立了 分化细胞系。 (4)重新定义组织中的顺式元件- 瞬时转染细胞特异性和免疫应答性 在目标#3中建立的路线。 特定核因子之间的相互作用 并将研究这些顺式元件。 成功完成 这些目标将大大增加我们对人类的理解。 调节类固醇生成中该关键酶的组织特异性表达。 成功完成本培训计划将使候选人能够 开始独立的研究生涯，研究分子 人类繁衍的基础