REGULATION OF COLLAGEN GENES COL4A5 AND COL4A6

胶原蛋白基因 COL4A5 和 COL4A6 的调控

• 批准号: 2391258
• 负责人: YOAV SEGAL
• 金额: $ 8.59万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-15 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2391258
• 关键词: DNA binding protein; DNA footprinting; RNase protection assay; basement membrane; collagen; gel mobility shift assay; genetic enhancer element; genetic mapping; genetic promoter element; genetic regulation; genetic regulatory element; genetic transcription; genetically modified animals; in situ hybridization; keratinocyte; laboratory mouse; northern blottings; posttranscriptional RNA processing; reporter genes; transcription factor; transfection; western blottings

The alpha5(IV) and alpha6(IV) chains of basement membrane type IV collagen are encoded by the genes COL4A5 and COL4A6, which are situated head-to- head on the X chromosome and divergently transcribed. Mutations in COL4A5 give rise to Alport syndrome (AS), an inherited disorder of glomerular basement membrane causing progressive renal insufficiency, often in association with hearing loss and ocular abnormalities. Deletions spanning the 5' termini of COL4A5 and COL4A6 have been identified in cases of AS-diffuse leiomyomatosis, in which AS is accompanied by disseminated smooth muscle tumors. The overall objective of this project is to elucidate the transcriptional regulation of COL4A5 and COL4A6. Specific aims are to map precisely and characterize the putative bidirectional promoter for these genes, and to define cis- and trans-acting regulatory elements which account for tissue- specific patterns of expression. The work will be performed in the outstanding environment of Harvard Medical School in the laboratories of Dr. Jing Zhou, an expert on the molecular genetics of basement membrane collagens, and Dr. William W. Chin, an expert on mammalian gene regulation. Consultants include Dr. Bjorn R. Olsen, an authority on collagens and extracellular matrix; Dr. Yoshihiko Yamada, an expert on the transcriptional regulation of type IV collagen genes; and Dr. Lawrence F. Brown, who has extensive experience with in situ hybridization. Methodologies to be used include RNase protection assays and primer extension analysis for identification of transcription start sites; transient transfections with reporter gene constructs for localization of promoter and cis-acting regulatory elements; development of transgenic mice to study tissue-specific gene expression; characterization of DNA-binding elements; and in situ hybridization. The applicant is a graduate of the Medical Scientist Training Program at Washington University, who has a record of accomplishment in the area of epithelial ion channels. He is committed to a basic scientific career in an academic division of nephrology. The proposed project will be carried out in a superb training environment with the strong support of experienced researchers. It will pose a series of strategic challenges which will provide excellent background for launching an independent career.

基底膜IV型胶原的α5(IV)和α6(IV)链 是由COL4A5和COL4A6基因编码的，这两个基因位于头对头之间 头部在X染色体上，并分化转录。COL4A5基因突变 导致Alport综合征(AS)，一种遗传性肾小球疾病 基底膜导致进行性肾功能不全，常见于 与听力损失和眼睛异常有关。删除部分 已在病例中发现跨越COL4A5和COL4A6 5‘末端的病例 AS弥漫性子宫肌瘤病，AS伴发播散性 平滑肌瘤。 这个项目的总体目标是阐明转录 COL4A5和COL4A6的调控。具体目标是准确地绘制和 鉴定这些基因的假定双向启动子，并 定义顺式和反式作用的调节元件，它们解释了组织- 特定的表达方式。 这项工作将在哈佛大学出色的环境中进行 医学专家周静博士的实验室里的医学院 基底膜胶原蛋白的分子遗传学和William W. 哺乳动物基因调控方面的专家Chin说道。顾问包括Dr。 比约恩·R·奥尔森是胶原蛋白和细胞外基质方面的权威； Yoshihiko Yamada，IV型转录调控专家 胶原基因；以及劳伦斯·F·布朗博士，他有丰富的经验 用原位杂交技术。要使用的方法包括RNase 保护性试验和引物延伸分析在鉴定中的应用 转录起始点；报告基因的瞬时转染 启动子和顺式作用调控基因的本地化构建 用于研究组织特异性基因的转基因小鼠的发展 表达；DNA结合元件的特性；以及原位 杂交。 申请人毕业于医学科学家培训计划，毕业于 华盛顿大学，他在以下领域取得了成就 上皮离子通道。他致力于基本的科学事业， 肾脏病学的一个学术分支。拟议的项目将得到实施 在一流的训练环境下，得到了 经验丰富的研究人员。这将带来一系列战略挑战 这将为推出独立的 职业生涯。