MOLECULAR GENETIC STUDIES OF C ELEGANS MORPHOGENESIS

多细胞生物的形态发生需要合成和 组装复杂的细胞外结构。 尽管几 细胞外蛋白质已被确定，很少有人知道如何 这些蛋白质的功能是产生有组织的大分子 结构. 本提案的长期目标是了解如何 细胞外蛋白质组装成大分子复合物以及它们如何组装 复合体参与生物体的形态发生和功能。 胶原蛋白是最丰富和普遍存在的细胞外 proteins. 线虫C.优雅， 已被证明会导致机体功能的严重缺陷， 形态学 强大的基因和分子分析， in C.秀丽隐杆线虫及其简单的解剖结构使其成为一个极好的模型系统 用于研究胶原蛋白的功能。 几个基因的突变， 编码形成角质层的胶原蛋白， C.基底膜胶原已经证明了秀丽线虫可以改变 生物体的形态和正常功能。 中的不同突变 这些胶原蛋白基因具有不同的遗传特性， 不同的形态缺陷。 额外的突变等位基因将是 产生的胶原蛋白的遗传和分子特性 可以检查缺陷。 将分析定义的突变的影响 通过创建携带体外诱变胶原的转基因菌株， 基因. 将生产和利用对胶原蛋白特异的抗体 为了分析将野生型和突变型胶原组装成 角质层和基底膜。 突变体结构的改变 通过电子显微镜分析角膜和基底膜， 利用特异性抗血清。 角质层胶原蛋白基因 显示出与其他影响生物体的基因相互作用 形态学 这些相互作用的基因可能编码蛋白质， 身体上的相互作用，所以他们也将受到进一步的遗传和 分子表征 这些研究将提供深入了解 胶原蛋白的遗传和分子特性及其在 生物体的形态发生和正常功能。 的接收

Morphogenesis of multicellular organisms requires the synthesis and assembly of complex extracellular structures. Although several extracellular proteins have been identified, little is known about how these proteins actually function to produce organized macromolecular structures. The long-term goals of this proposal are to understand how extracellular proteins assemble into macromolecular complexes and how these complexes participate in the morphogenesis and functioning of the organism. Collagens are the most abundant and ubiquitous of the extracellular proteins. Mutations in several collagen genes in the nematode, C. elegans, have been shown to cause severe defects in organismal function and morphology. The powerful genetic and molecular analyses that are possible in C. elegans, and its simple anatomy, make it an excellent model system for studying the functions of collagens. Mutations in several genes that encode collagens that form the cuticle and in one gene that encodes a basement membrane collagen of C. elegans have been shown to alter the morphology and normal functioning of the organism. Different mutations in these collagen genes have different genetic properties and can cause very different morphological defects. Additional mutant alleles will be generated so that the genetic and molecular properties of further collagen defects can be examined. The effects of defined mutations will be analyzed by creating transgenic strains carrying in vitro mutagenized collagen genes. Antibodies specific for the collagens will be produced and utilized to analyze the steps of assembly of wild-type and mutant collagens into the cuticle and basement membranes. Alterations in the structure of mutant cuticles and basement membranes will be analyzed by electronmicroscopy, utilizing the specific antisera. The cuticle collagen genes have been shown to interact genetically with other genes that affect organismal morphology. These interacting genes are likely to encode proteins that interact physically, so they will also be subjected to further genetic and molecular characterization. These studies will provide insights into the genetic and molecular properties of collagens and their roles in the morphogenesis and normal functioning of the organism. The receipt of an