STRUCTURE AND FUNCTION OF THE THROMBOSPONDIN GENE FAMILY

血小板反应蛋白基因家族的结构和功能

• 批准号: 6296254
• 负责人: PAUL BORNSTEIN
• 金额: $ 10.57万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6296254
• 关键词: autosomal dominant trait; cytokine; extracellular matrix; gene deletion mutation; gene expression; genetic promoter element; genetically modified animals; glycoprotein structure; growth factor; guinea pigs; laboratory mouse; laboratory rabbit; messenger RNA; nucleic acid sequence; posttranscriptional RNA processing; protein structure function; thrombospondins; tissue /cell culture

Extracellular matrix proteins are known to serve not only as structural elements in tissues but also to influence cell shape, movement, biosynthetic phenotype, and proliferation. In the animal, these functions contribute to embryonic development, morphogenesis, and tissue differentiation. Thrombospondin (TSP) belongs to a class of extracellular proteins that is thought to modulate cell-matrix interactions, and may play a role in regulating cell attachment, proliferation, and migration, and in angiogenesis. This laboratory has demonstrated that there are at least three expressed genes coding for related TSPs. The existence of three protein chains that are likely to demonstrate both similarities and crucial differences in structure and function obliges a reevaluation of previous work relating to the location and function of TSP, and greatly increases the potential complexity of TSP-cell interactions and the range of functions which this group of proteins could perform. Our objectives in this proposal are to isolate and characterize the three mouse TSP genes, to compare the pattern of mRNA and protein expression in mouse cells and tissues, and to assess the response of these genes to growth factors and to cytokines. Since mutations in TSPs are likely to exert dominant negative effects, minigenes that exclude several exons will be constructed and introduced into mice. Transgenic mice carrying such genes could demonstrate a phenotype that reflects the function of the mutated TSP. Finally, targeted disruption of the TSP genes will be performed by homologous recombination in somatic cells to evaluate the requirement of the TSPs for cell attachment, migration, and proliferation. It is expected that these experiments will clarify the function of the individual TSPs and address the possibility of the existence of heterotrimeric TSPs. It will be instructive to examine the regulation of the three genes and to relate these findings to the pattern of expression and function of the three proteins. This information should increase our understanding of the role of the TSPs in the response of tissues to injury and, in particular, to the reaction of blood vessel wall to endothelial damage.

已知细胞外基质蛋白不仅作为结构蛋白， 组织中的元素，也影响细胞的形状，运动， 生物合成表型和增殖。 在动物中，这些功能 有助于胚胎发育、形态发生和组织 分化 血小板反应蛋白（TSP）属于一类细胞外 蛋白质被认为是调节细胞基质相互作用，并可能发挥作用， 在调节细胞附着、增殖和迁移中的作用， 血管生成 该实验室已经证明，至少有三种表达 编码相关TSP的基因。 三条蛋白质链的存在 很可能在以下方面既有相似之处， 结构和功能要求重新评价以前的工作， TSP的定位和作用，大大提高了 TSP-细胞相互作用的复杂性和这一作用的功能范围 一组蛋白质可以执行。 我们在这项建议中的目标是 分离和鉴定三种小鼠TSP基因，比较其模式， mRNA和蛋白质在小鼠细胞和组织中的表达，并评估 这些基因对生长因子和细胞因子的反应。 以来 TSPs的突变可能会产生显性负效应， 将构建排除几个外显子的基因组并导入小鼠。 携带这些基因的转基因小鼠可以表现出一种表型， 反映了突变TSP的功能。 最后，有针对性地破坏 TSP基因将通过体细胞中的同源重组进行， 细胞以评估TSP对细胞附着的需求， 移民和扩散。 预计这些实验将阐明 个别TSP，并解决存在的可能性， 异源三聚体TSP。 审查对下列方面的管制将是有益的： 这三个基因，并将这些发现与表达模式联系起来， 三种蛋白质的功能。 这些信息将增加我们的 了解TSP在组织对损伤的反应中的作用 特别是血管壁对内皮细胞的反应 损害