GENETIC STUDIES OF NON-ERYTHROID SPECTRIN

血影蛋白的功能将在一个模型系统中进行分析，果蝇 黑猩猩。血影蛋白是一种膜相关的细胞骨架蛋白 这在确定红细胞形状方面起着关键作用。 在非红系细胞中，血影蛋白也扮演着未知但必不可少的角色。 由于我们最近的工作已经发现了阿尔法-幽灵蛋白 变种人，我们处于独特的地位，能够直接评估 血影蛋白在非红系细胞中的功能。 使用生化、分子遗传学和经典的组合 基因分析，我们将检验有关血影蛋白功能的各种假说 在细胞、发育和组织的非红系组织中 水平。将产生β-幽灵蛋白突变体，这些突变体与 最近发现的阿尔法血影蛋白突变体，将被用于 研究血影蛋白在体内的功能。表型分析将决定 镶嵌，镶嵌完全缺乏功能的血影蛋白的影响；镶嵌 分析将确定血影蛋白缺失在选定的 发育阶段。这些表型信息将被用来设计 用光影蛋白-DNA构建拯救零突变体的实验 将测试特定方面的细胞或发育作用 血影蛋白组织，如膜结合，或血影蛋白链- 长度。 尽管几乎所有的人类细胞中都存在血影蛋白亚型，但 幽灵蛋白缺陷对神经系统和全身健康的影响 在非红系细胞中的表达才刚刚开始探索。这个 拟议的实验将提供对血影蛋白的深入了解 在非红系细胞中的作用。

Spectrin function will be analyzed in a model system, Drosophila melanogaster. Spectrin is a membrane-associated cytoskeletal protein that plays a key role in the determination of erythrocyte cell shape. spectrin also plays unknown, but essential, roles in non-erythroid cells. As a result of our recent work that has identified alpha-spectrin mutants, we are in the singular position of being able to directly assess the functions of spectrin in non-erythroid cells. Using a combination of biochemical, molecular genetic, and classical genetic analyses, we will test various hypotheses for spectrin function in non-erythroid tissue at the cellular, developmental and organismal level. Beta-spectrin mutants will be produced and these, together with recently identified alpha-spectrin mutants, will be utilized to investigate spectrin function in vivo. Phenotype analysis will determine the effects of the complete absence of functional spectrin; mosaic analysis will determine the effects of spectrin absence at selected developmental stages. This phenotype information will be used to design experiments that rescue null mutants with spectrin-DNA constructs that will test the cellular or developmental role of a specific aspect of spectrin organization, such as membrane binding, or spectrin chain- length. Although spectrin isoforms are present in nearly all human cells, the neurological and general health consequences of defective spectrin expression in non-erythroid cells are only beginning to be explored. The proposed experiments will provide an in-depth understanding of spectrin's role in non-erythroid cells.