STRUCTURE & REGULATION OF BASEMENT MEMBRANE PROTEOGLYCAN

结构

• 批准号: 3304842
• 负责人: JOHN ROBERT HASSELL
• 金额: $ 19.1万
• 依托单位: EYE AND EAR INSTITUTE OF PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3304842
• 关键词: RNA splicing; antibody formation; basement membrane; cell differentiation; complementary DNA; computer assisted sequence analysis; extracellular matrix; gel electrophoresis; gene expression; glycoprotein structure; heparan sulfate; laboratory mouse; laboratory rabbit; membrane structure; messenger RNA; molecular cloning; protein structure function; proteoglycan; transcription factor

Basement membranes are thin sheets of extracellular matrix that separate dissimilar cells and tissues. This matrix is difficult to isolate from native tissues but tumor cell lines which exclusively produce basement membrane have proven useful as a model system for identifying and characterizing the major components of this type of extracellular matrix. A heparan sulfate containing proteoglycan has been isolated and partially characterized using these model cell lines. Its core protein is an exceptionally large gene product (400,00 Mr) containing homologies to N- cam, laminin and low density lipoprotein receptor in the regions os far cloned. This proteoglycan, or its counterpart in native and cell attachment. The long term objective of this proposal is to establish the complete structure of the proteoglycan and identify the process involved in the regulation of its production. The specific aims are: (1) to complete the characterization of the proteoglycan core protein gene product by constructing cDNA libraries and, using existing clones, isolate the cDNA clones needed to deduce the entire core protein structure; (2) to characterize the proteoglycan gene by using the cDNA clones to isolate genomic clones, to determine the intron/exon structure and to identify the cis-acting region of the gene which controls transcription; (3) to establish the regulation of proteoglycan gene expression during differentiation of F9 cells by using the cDNA clones to measure the kinetics of change of mRNA levels and mRNA synthesis and mRNA levels upon response to exogenous matrix. The results of these studies will provide an understanding of the proteoglycan core protein structure and the mechanisms involved in its production during the biogenesis and maintenance of basement membranes.

地下膜是细胞外基质的薄片 不同的细胞和组织。 这个矩阵很难与 天然组织，但肿瘤细胞系只产生地下室 事实证明，膜是识别和 表征这种类型的细胞外基质的主要组成部分。 已经分离出含有蛋白聚糖的乙酰乙酰硫酸乙酰肝素 使用这些模型细胞系进行了特征。 它的核心蛋白是 具有非基因产物（400,00 MR），其中含有N-的同源物 CAM，层粘连蛋白和低密度脂蛋白受体在远处OS 克隆。 该蛋白聚糖或本机和细胞中的对应物 依恋。 该提议的长期目标是建立 蛋白聚糖的完整结构并确定所涉及的过程 其生产的调节。 具体目的是：（1）完成 蛋白聚糖核心蛋白基因产物的表征 构建cDNA库，并使用现有克隆分离cDNA 克隆需要推断整个核心蛋白质结构； （2）至 通过使用cDNA克隆分离蛋白聚糖基因表征蛋白聚糖基因 基因组克隆，确定内含子/外显子结构并确定 控制转录的基因的顺式作用区域； （3）到 建立蛋白聚糖基因表达的调节 通过使用cDNA克隆测量F9细胞的分化 mRNA水平变化和mRNA合成和mRNA水平的动力学 对外源基质的响应。 这些研究的结果将提供 了解蛋白聚糖核心蛋白结构和机制 在生物发生和维持期间参与其生产 地下室膜。