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。尽管有几个 胞外蛋白已被鉴定，但对它是如何形成的知之甚少 这些蛋白质实际上能产生有组织的大分子。 结构。这项提案的长期目标是了解如何 胞外蛋白质组装成大分子复合体以及这些 复合体参与生物体的形态发生和功能。 胶原蛋白是细胞外最丰富、最普遍的物质 蛋白质。线虫线虫中几个胶原基因的突变， 已被证明会导致严重的器官功能缺陷 形态学。强大的遗传和分子分析 在线虫中，以及它简单的解剖结构，使它成为一个很好的模型系统 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，并在一个编码 线虫的基底膜胶原蛋白已被证明可以改变 生物体的形态和正常功能。基因中的不同突变 这些胶原蛋白基因具有不同的遗传特性，可以导致非常 不同的形态缺陷。其他突变的等位基因将是 因此进一步的胶原蛋白的遗传和分子性质 缺陷是可以检查的。我们将分析已定义的突变的影响 通过创造携带体外诱变胶原的转基因菌株 基因。将产生和利用针对胶原蛋白的特异性抗体 分析野生型和突变型胶原蛋白组装成 角质层和基底膜。突变体的结构变化 角质层和基底膜将通过电子显微镜进行分析， 利用特定的抗血清。角质层胶原蛋白基因已经 显示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很可能编码的蛋白质 身体上的相互作用，所以他们还会受到进一步的遗传和 分子表征。这些研究将为我们提供对 胶原蛋白的遗传和分子特性及其在糖尿病发病中的作用 生物体的形态发生和正常功能。收到一封信

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