REGULATION OF COLLAGEN GENES COL4A5 AND COL4A6

胶原蛋白基因 COL4A5 和 COL4A6 的调控

• 批准号: 2900076
• 负责人: YOAV SEGAL
• 金额: $ 3.47万
• 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 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2900076
• 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α 5（IV）和α 6（IV）链 是由基因COL 4A 5和COL 4A 6编码的，它们位于头对- 头部在X染色体上，并被发散地转录。 COL 4A 5突变 导致Alport综合征（AS），一种遗传性肾小球疾病， 基底膜引起进行性肾功能不全，通常在 与听力损失和眼部异常有关。 缺失 跨越COL 4A 5和COL 4A 6的5'末端的蛋白质已经在一些病例中被鉴定， AS-弥漫性平滑肌瘤病，其中AS伴有弥漫性 平滑肌肿瘤 这个项目的总体目标是阐明转录 COL 4A 5和COL 4A 6的调节。 具体目标是精确绘制地图， 表征这些基因的推定双向启动子，并 定义顺式和反式作用的调节元件，其解释组织- 具体的表达方式。 这项工作将在哈佛杰出的环境中进行 医学院周敬博士的实验室， 基底膜胶原蛋白的分子遗传学，威廉W。 他是哺乳动物基因调控方面的专家。 顾问包括博士。 比约恩河奥尔森博士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;和Dr. Lawrence F.布朗拥有丰富的经验， 用原位杂交技术。 使用的方法包括RNase 保护测定和引物延伸分析，用于鉴定 转录起始位点;报告基因瞬时转染 用于定位启动子和顺式作用调节因子的构建体 元件;研究组织特异性基因的转基因小鼠的开发 表达; DNA结合元件的表征;和原位 杂交方法 申请人是医学科学家培训计划的毕业生， 华盛顿大学，他在该领域的成就记录， 上皮细胞离子通道 他致力于基础科学事业， 肾脏学的一个学术分支。 拟议项目将在 在一流的训练环境中， 经验丰富的研究人员。 将带来一系列战略挑战 这将为启动一个独立的 事业